Method for Increasing the Vigor and/or Crop Yield of Agricultural Plants Under Essentially Non-Existent Pathogen Pressure

ABSTRACT

A method for increasing the vigor and/or crop yield of agricultural plants under essentially non-existent pathogen pressure, wherein the plants, the plant propagules, the seed of the plants and/or the locus where the plants are growing or are intended to grow are treated with an effective amount of a composition comprising a) the  Bacillus subtilis  strain with NRRL Accession No. B-21661 or a cell-free extract thereof, and/or a mutant of this strain or extract having all the identifying characteristics of the respective strain or extract as component (I), and b) optionally at least one chemical compound as component (II), selected from the active compound groups A) to J): A) strobilurins; B) carboxamides; C) azoles; D) heterocyclic compounds; E) carbamates; F) other active substances; G) growth regulators; H) herbicides; J) insecticides.

The present invention relates to a method for increasing the vigorand/or crop yield of agricultural plants under essentially non-existentpathogen pressure, wherein the plants, the plant propagules, the seed ofthe plants and/or the locus where the plants are growing or are intendedto grow are treated with an effective amount of a composition comprising

-   -   a) the Bacillus subtilis strain with NRRL Accession No. B-21661        or a cell-free extract thereof, and/or a mutant of this strain        or extract having all the identifying characteristics of the        respective strain or extract as component (I), and    -   b) optionally at least one chemical compound as component (II),        selected from the active compound groups A) to J):        -   A) strobilurins selected from the group consisting of            azoxystrobin, dimoxystrobin, enestroburin, fluoxastrobin,            kresoxim-methyl, metominostrobin, orysastrobin,            picoxystrobin, pyraclostrobin, pyribencarb, trifloxystrobin,            2-(2-(6-(3-chloro-2-methyl-phenoxy)-5-fluoro-pyrimidin-4-yloxy)-phenyl)-2-methoxyimino-N-methyl-acetamide,            3-methoxy-2-(2-(N-(4-methoxy-phenyl)-cyclopropane-carboximidoylsulfanylmethyl)-phenyl)-acrylic            acid methyl ester,            methyl(2-chloro-5-[1-(3-methylbenzyloxyimino)-ethyl]benzyl)carbamate            and            2-(2-(3-(2,6-dichlorophenyl)-1-methyl-allylideneaminooxymethyl)-phenyl)-2-methoxyimino-N-methyl-acetamide;        -   B) carboxamides selected from the group consisting of            -   carboxanilides: benalaxyl, benalaxyl-M, benodanil,                bixafen, boscalid, carboxin, fenfuram, fenhexamid,                flutolanil, furametpyr, isopyrazam, isotianil,                kiralaxyl, mepronil, metalaxyl, metalaxyl-M (mefenoxam),                ofurace, oxadixyl, oxycarboxin, penthiopyrad, sedaxane,                tecloftalam, thifluzamide, tiadinil,                2-amino-4-methyl-thiazole-5-carboxanilide,                2-chloro-N-(1,1,3-trimethyl-indan-4-yl)-nicotinamide,                N-(3′,4′,5′-trifluorobiphenyl-2-yl)-3-difluoromethyl-1-methyl-1H-pyrazole-4-carboxamide,                N-(4′-trifluoromethylthiobiphenyl-2-yl)-3-difluoromethyl-1-methyl-1H-pyrazole-4-carboxamide,                N-(2-(1,3-dimethyl-butyl)-phenyl)-1,3-dimethyl-5-fluoro-1H-pyrazole-4-carboxamide                and                N-(2-(1,3,3-trimethyl-butyl)-phenyl)-1,3-dimethyl-5-fluoro-1H-pyrazole-4-carboxamide;            -   carboxylic morpholides: dimethomorph, flumorph,                pyrimorph;            -   benzoic acid amides: flumetover, fluopicolide,                fluopyram, zoxamide,                N-(3-Ethyl-3,5,5-trimethyl-cyclohexyl)-3-formylamino-2-hydroxy-benzamide;            -   other carboxamides: carpropamid, dicyclomet,                mandiproamid, oxytetracyclin, silthiofarm and                N-(6-methoxy-pyridin-3-yl)cyclopropanecarboxylic acid                amide;        -   C) azoles selected from the group consisting of            -   triazoles: azaconazole, bitertanol, bromuconazole,                cyproconazole, difenoconazole, diniconazole,                diniconazole-M, epoxiconazole, fenbuconazole,                fluquinconazole, flusilazole, flutriafol, hexaconazole,                imibenconazole, ipconazole, metconazole, myclobutanil,                oxpoconazole, paclobutrazole, penconazole,                propiconazole, prothioconazole, simeconazole,                tebuconazole, tetraconazole, triadimefon, triadimenol,                triticonazole, uniconazole,                1-(4-chloro-phenyl)-2-([1,2,4]triazol-1-yl)-cycloheptanol;            -   imidazoles: cyazofamid, imazalil, pefurazoate,                prochloraz, triflumizol;            -   benzimidazoles: benomyl, carbendazim, fuberidazole,                thiabendazole;            -   others: ethaboxam, etridiazole, hymexazole and                2-(4-chloro-phenyl)-N-[4-(3,4-dimethoxy-phenyl)-isoxazol-5-yl]-2-prop-2-ynyloxy-acetamide;        -   D) heterocyclic compounds selected from the group consisting            of            -   pyridines: fluazinam, pyrifenox,                3-[5-(4-chloro-phenyl)-2,3-dimethyl-isoxazolidin-3-yl]-pyridine,                3-[5-(4-methyl-phenyl)-2,3-dimethyl-isoxazolidin-3-yl]-pyridine,                2,3,5,6-tetra-chloro-4-methanesulfonyl-pyridine,                3,4,5-trichloropyridine-2,6-di-carbonitrile,                N-(1-(5-bromo-3-chloro-pyridin-2-yl)-ethyl)-2,4-dichloronicotinamide,                N-[(5-bromo-3-chloro-pyridin-2-yl)-methyl]-2,4-dichloro-nicotinamide;            -   pyrimidines: bupirimate, cyprodinil, diflumetorim,                fenarimol, ferimzone, mepanipyrim, nitrapyrin, nuarimol,                pyrimethanil;            -   piperazines: triforine;            -   pyrroles: fenpiclonil, fludioxonil;            -   morpholines: aldimorph, dodemorph, dodemorph-acetate,                fenpropimorph, tridemorph;            -   piperidines: fenpropidin;            -   dicarboximides: fluoroimid, iprodione, procymidone,                vinclozolin;            -   non-aromatic 5-membered heterocycles: famoxadone,                fenamidone, flutianil, octhilinone, probenazole,                5-amino-2-isopropyl-3-oxo-4-ortho-tolyl-2,3-dihydro-pyrazole-1-carbothioic                acid S-allyl ester;            -   others: acibenzolar-S-methyl, amisulbrom, anilazin,                blasticidin-S, captafol, captan, chinomethionat,                dazomet, debacarb, diclomezine, difenzoquat,                difenzoquat-methylsulfate, fenoxanil, Folpet, oxolinic                acid, piperalin, proquinazid, pyroquilon, quinoxyfen,                triazoxide, tricyclazole,                2-butoxy-6-iodo-3-propylchromen-4-one,                5-chloro-1-(4,6-dimethoxy-pyrimidin-2-yl)-2-methyl-1H-benzoimidazole,                5-chloro-7-(4-methyl-piperidin-1-yl)-6-(2,4,6-trifluorophenyl)-[1,2,4]triazolo[1,5-a]pyrimidine                and                5-ethyl-6-octyl-[1,2,4]triazolo[1,5-a]pyrimidine-7-ylamine;        -   E) carbamates selected from the group consisting of            -   thio- and dithiocarbamates: ferbam, mancozeb, maneb,                metam, methasulphocarb, metiram, propineb, thiram,                zineb, ziram;            -   carbamates: benthiavalicarb, diethofencarb,                iprovalicarb, propamocarb, propamocarb hydrochlorid,                valiphenal and                N-(1-(1-(4-cyano-phenyl)-ethanesulfonyl)-but-2-yl)                carbamic acid-(4-fluorophenyl) ester;        -   F) other active substances selected from the group            consisting of            -   guanidines: guanidine, dodine, dodine free base,                guazatine, guazatine-acetate, iminoctadine,                iminoctadine-triacetate, iminoctadine-tris(albesilate);            -   antibiotics: kasugamycin, kasugamycin                hydrochloride-hydrate, streptomycin, polyoxine,                validamycin A, streptomycin;            -   nitrophenyl derivates: binapacryl, dinobuton, dinocap,                nitrthal-isopropyl, tecnazen,            -   organometal compounds: fentin salts, such as                fentin-acetate, fentin chloride or fentin hydroxide;            -   sulfur-containing heterocyclyl compounds: dithianon,                isoprothiolane;            -   organophosphorus compounds: edifenphos, fosetyl,                fosetyl-aluminum, iprobenfos, phosphorous acid and its                salts, pyrazophos, tolclofos-methyl;            -   organochlorine compounds: chlorothalonil, dichlofluanid,                dichlorophen, flusulfamide, hexachlorobenzene,                pencycuron, pentachlorphenole and its salts, phthalide,                quintozene, thiophanate-methyl, tolylfluanid,                N-(4-chloro-2-nitro-phenyl)-N-ethyl-4-methyl-benzenesulfonamide;            -   inorganic active substances: Bordeaux mixture, copper                acetate, copper hydroxide, copper oxychloride, basic                copper sulfate, sulfur;            -   others: biphenyl, bronopol, cyflufenamid, cymoxanil,                diphenylamin, metrafenone, mildiomycin, oxin-copper,                prohexadione-calcium, spirox-amine, tolylfluanid,                N-(cyclopropylmethoxyimino-(6-difluoro-methoxy-2,3-difluoro-phenyl)-methyl)-2-phenyl                acetamide,                N′-(4-(4-chloro-3-trifluoromethyl-phenoxy)-2,5-dimethyl-phenyl)-N-ethyl-N-methyl                formamidine,                N′-(4-(4-fluoro-3-trifluoronnethyl-phenoxy)-2,5-dinnethyl-phenyl)-N-ethyl-N-methyl                formamidine,                N′-(2-methyl-5-trifluoromethyl-4-(3-trimethylsilanyl-propoxy)-phenyl)-N-ethyl-N-methyl                formamidine,                N′-(5-difluoromethyl-2-methyl-4-(3-trimethylsilanyl-propoxy)-phenyl)-N-ethyl-N-methyl                formamidine,                2-{1-[2-(5-methyl-3-trifluoromethyl-pyrazole-1-yl)-acetyl]-piperidin-4-yl}-thiazole-4-carboxylic                acid methyl-(1,2,3,4-tetrahydro-naphthalen-1-yl)-amide,                2-{142-(5-methyl-3-trifluoromethyl-pyrazole-1-yl)-acetyl]-piperidin-4-yll-thiazole-4-carboxylic                acid                methyl-(R)-1,2,3,4-tetrahydro-naphthalen-1-yl-amide,                acetic acid                6-tert.-butyl-8-fluoro-2,3-dimethyl-quinolin-4-yl ester                and methoxy-acetic acid                6-tert-butyl-8-fluoro-2,3-dimethyl-quinolin-4-yl ester;                fentin acetate, fentin chloride, fentin hydroxide;        -   G) plant growth regulators (PGRs) selected from the group            consisting of abscisic acid, amidochlor, ancymidol,            6-benzylaminopurine, brassinolide, butralin, chlormequat            (chlormequat chloride), choline chloride, cyclanilide,            daminozide, dikegulac, dimethipin, 2,6-dimethylpuridine,            ethephon, flumetralin, flurprimidol, fluthiacet,            forchlorfenuron, gibberellic acid, inabenfide,            indole-3-acetic acid, maleic hydrazide, mefluidide, mepiquat            (mepiquat chloride), naphthaleneacetic acid,            N-6-benzyladenine, paclobutrazol, prohexadione            (prohexadione-calcium), prohydrojasmon, thidiazuron,            triapenthenol, tributyl phosphorotrithioate,            2,3,5-tri-iodobenzoic acid, trinexapac-ethyl and            uniconazole;        -   H) herbicides selected from the group consisting of            -   acetamides: acetochlor, alachlor, butachlor,                dimethachlor, dimethenamid, flufenacet, mefenacet,                metolachlor, metazachlor, napropamide, naproanilide,                pethoxamid, pretilachlor, propachlor, thenylchlor;            -   amino acid derivatives: bilanafos, glufosinate,                sulfosate;            -   aryloxyphenoxypropionates: clodinafop, cyhalofop-butyl,                fenoxaprop, fluazifop, haloxyfop, metamifop,                propaquizafop, quizalofop, quizalofop-P-tefuryl;            -   Bipyridyls: diquat, paraquat;            -   (thio)carbamates: asulam, butylate, carbetamide,                desmedipham, dimepiperate, eptam (EPTC), esprocarb,                molinate, orbencarb, phenmedipham, prosulfocarb,                pyributicarb, thiobencarb, triallate;            -   cyclohexanediones: butroxydim, clethodim, cycloxydim,                profoxydim, sethoxydim, tepraloxydim, tralkoxydim;            -   dinitroanilines: benfluralin, ethalfluralin, oryzalin,                pendimethalin, prodiamine, trifluralin;            -   diphenyl ethers: acifluorfen, aclonifen, bifenox,                diclofop, ethoxyfen, fomesafen, lactofen, oxyfluorfen;            -   hydroxybenzonitriles: bomoxynil, dichlobenil, ioxynil;            -   imidazolinones: imazamethabenz, imazamox, imazapic,                imazapyr, imazaquin, imazethapyr;            -   phenoxy acetic acids: clomeprop,                2,4-dichlorophenoxyacetic acid (2,4-D), 2,4-DB,                dichlorprop, MCPA, MCPA-thioethyl, MCPB, Mecoprop;            -   pyrazines: chloridazon, flufenpyr-ethyl, fluthiacet,                norflurazon, pyridate;            -   pyridines: aminopyralid, clopyralid, diflufenican,                dithiopyr, fluridone, fluroxypyr, picloram, picolinafen,                thiazopyr;            -   sulfonyl ureas: amidosulfuron, azimsulfuron,                bensulfuron, chlorimuron-ethyl, chlorsulfuron,                cinosulfuron, cyclosulfamuron, ethoxysulfuron,                flazasulfuron, flucetosulfuron, flupyrsulfuron,                foramsulfuron, halosulfuron, imazosulfuron,                iodosulfuron, mesosulfuron, metsulfuron-methyl,                nicosulfuron, oxasulfuron, primisulfuron, prosulfuron,                pyrazosulfuron, rimsulfuron, sulfometuron,                sulfosulfuron, thifensulfuron, triasulfuron, tribenuron,                trifloxysulfuron, triflusulfuron, tritosulfuron,                1-((2-chloro-6-propylimidazo[1,2-b]pyridazin-3-yl)sulfonyl)-3-(4,6-dimethoxy-pyrimidin-2-yl)urea;            -   triazines: ametryn, atrazine, cyanazine, dimethametryn,                ethiozin, hexazinone, metamitron, metribuzin, prometryn,                simazine, terbuthylazine, terbutryn, triaziflam;            -   ureas: chlorotoluron, daimuron, diuron, fluometuron,                isoproturon, linuron, methabenzthiazuron, tebuthiuron;            -   other acetolactate synthase inhibitors:                bispyribac-sodium, cloransulam-methyl, diclosulam,                florasulam, flucarbazone, flumetsulam, metosulam,                ortho-sulfamuron, penoxsulam, propoxycarbazone,                pyribambenz-propyl, pyribenzoxim, pyriftalid,                pyriminobac-methyl, pyrimisulfan, pyrithiobac,                pyroxasulfone, pyroxsulam;            -   others: amicarbazone, aminotriazole, anilofos,                beflubutamid, benazolin, bencarbazone, benfluresate,                benzofenap, bentazone, benzobicyclon, bromacil,                bromobutide, butafenacil, butamifos, cafenstrole,                carfentrazone, cinidon-ethlyl, chlorthal, cinmethylin,                clomazone, cumyluron, cyprosulfamide, dicamba,                difenzoquat, diflufenzopyr, Drechslera monoceras,                endothal, ethofumesate, etobenzanid, fentrazamide,                flumiclorac-pentyl, flumioxazin, flupoxam,                flurochloridone, flurtamone, indanofan, isoxaben,                isoxaflutole, lenacil, propanil, propyzamide,                quinclorac, quinmerac, mesotrione, methyl arsonic acid,                naptalam, oxadiargyl, oxadiazon, oxaziclomefone,                pentoxazone, pinoxaden, pyraclonil, pyraflufen-ethyl,                pyrasulfotole, pyrazoxyfen, pyrazolynate, quinoclamine,                saflufenacil, sulcotrione, sulfentrazone, terbacil,                tefuryltrione, tembotrione, thiencarbazone, topramezone,                4-hydroxy-3-[2-(2-methoxy-ethoxymethyl)-6-trifluoromethyl-pyridine-3-carbonyl]-bicyclo[3.2.1]oct-3-en-2-one,                (3-[2-chloro-4-fluoro-5-(3-methyl-2,6-dioxo-4-trifluoromethyl-3,6-dihydro-2H-pyrimidin-1-yl)-phenoxy]-pyridin-2-yloxy)-acetic                acid ethyl ester,                6-amino-5-chloro-2-cyclopropyl-pyrimidine-4-carboxylic                acid methyl ester,                6-chloro-3-(2-cyclopropyl-6-methyl-phenoxy)-pyridazin-4-ol,                4-amino-3-chloro-6-(4-chloro-phenyl)-5-fluoro-pyridine-2-carboxylic                acid,                4-amino-3-chloro-6-(4-chloro-2-fluoro-3-methoxy-phenyl)-pyridine-2-carboxylic                acid methyl ester, and                4-amino-3-chloro-6-(4-chloro-3-dimethylamino-2-fluoro-phenyl)-pyridine-2-carboxylic                acid methyl ester;        -   J) insecticides selected from the group consisting of            -   organo(thio)phosphates: acephate, azamethiphos,                azinphos-methyl, chlorpyrifos, chlorpyrifos-methyl,                chlorfenvinphos, diazinon, dichlorvos, dicrotophos,                dimethoate, disulfoton, ethion, fenitrothion, fenthion,                isoxathion, malathion, methamidophos, methidathion,                methyl-parathion, mevinphos, monocrotophos,                oxydemeton-methyl, paraoxon, parathion, phenthoate,                phosalone, phosmet, phosphamidon, phorate, phoxim,                pirimiphos-methyl, profenofos, prothiofos, sulprophos,                tetrachlorvinphos, terbufos, triazophos, trichlorfon;            -   carbamates: alanycarb, aldicarb, bendiocarb,                benfuracarb, carbaryl, carbofuran, carbosulfan,                fenoxycarb, furathiocarb, methiocarb, methomyl, oxamyl,                pirimicarb, propoxur, thiodicarb, triazamate;            -   pyrethroids: allethrin, bifenthrin, cyfluthrin,                cyhalothrin, cyphenothrin, cypermethrin,                alpha-cypermethrin, beta-cypermethrin,                zeta-cypermethrin, deltamethrin, esfenvalerate,                etofenprox, fenpropathrin, fenvalerate, imiprothrin,                lambda-cyhalothrin, permethrin, prallethrin, pyrethrin I                and II, resmethrin, silafluofen, tau-fluvalinate,                tefluthrin, tetramethrin, tralomethrin, transfluthrin,                profluthrin, dimefluthrin;            -   insect growth regulators: a) chitin synthesis                inhibitors: benzoylureas: chlorfluazuron, cyramazin,                diflubenzuron, flucycloxuron, flufenoxuron,                hexaflumuron, lufenuron, novaluron, teflubenzuron,                triflumuron; buprofezin, diofenolan, hexythiazox,                etoxazole, clofentazine; b) ecdysone antagonists:                halofenozide, methoxyfenozide, tebufenozide,                azadirachtin; c) juvenoids: pyriproxyfen, methoprene,                fenoxycarb; d) lipid biosynthesis inhibitors:                spirodiclofen, spiromesifen, spirotetramat;            -   nicotinic receptor agonists/antagonists compounds:                clothianidin, dinotefuran, imidacloprid, thiamethoxam,                nitenpyram, acetamiprid, thiacloprid,                1-(2-chloro-thiazol-5-ylmethyl)-2-nitrimino-3,5-dimethyl-[1,3,5]triazinane;            -   GABA antagonist compounds: endosulfan, ethiprole,                fipronil, vaniliprole, pyrafluprole, pyriprole,                5-amino-1-(2,6-dichloro-4-methyl-phenyl)-4-sulfinamoyl-1H-pyrazole-3-carbothioic                acid amide;            -   macrocyclic lactone insecticides: abamectin, emamectin,                milbemectin, lepimectin, spinosad, spinetoram;            -   mitochondrial electron transport inhibitor (METI) I                acaricides: fenazaquin, pyridaben, tebufenpyrad,                tolfenpyrad, flufenerim;            -   METI II and III compounds: acequinocyl, fluacyprim,                hydramethylnon;            -   uncouplers: chlorfenapyr;            -   oxidative phosphorylation inhibitors: cyhexatin,                diafenthiuron, fenbutatin oxide, propargite;            -   moulting disruptor compounds: cryomazine;            -   mixed function oxidase inhibitors: piperonyl butoxide;            -   sodium channel blockers: indoxacarb, metaflumizone;            -   others: benclothiaz, bifenazate, cartap, flonicamid,                pyridalyl, pymetrozine, sulfur, thiocyclam,                flubendiamide, chlorantraniliprole, cyazypyr (HGW86),                cyenopyrafen, flupyrazofos, cyflumetofen, amidoflumet,                imicyafos, bistrifluron, and pyrifluquinazon.

All mixtures set forth above are also an embodiment of the presentinvention.

In one embodiment, the invention relates to the use of the Bacillussubtilis strain with NRRL Accession No. B-21661 or a cell-free extractthereof, and/or a mutant of this strain or extract having all theidentifying characteristics of the respective strain or extract ascomponent (I), and optionally of at least one chemical compound ascomponent (II), selected from the active compound groups A) to J), forincreasing the vigor and/or crop yield of agricultural plants underessentially non-existent pathogen pressure.

The below remarks as to preferred embodiments of component (I) as wellas component (II) and respective mixtures and/or compositions comprisingcomponent (I) as well as component (II), to their preferred use andmethods of using them are to be understood either each on their own orpreferably in combination with each other.

In a preferred embodiment, the present invention relates to a method forincreasing the vigor and/or crop yield of agricultural plants underessentially non-existent pathogen pressure, wherein the plants, theplant propagules, the seed of the plants and/or the locus where theplants are growing or are intended to grow are treated with an effectiveamount of a composition comprising

-   -   a) the Bacillus subtilis strain with NRRL Accession No. B-21661        or a cell-free extract thereof, and/or a mutant of this strain        or extract having all the identifying characteristics of the        respective strain or extract as component (I), and    -   b) one chemical compound as component (II), selected from the        active compound groups A) to J) as defined above,        in a weight ratio of from 100:1 to 1:100.

In another preferred embodiment, the present invention relates to amethod for increasing the vigor and/or crop yield of agricultural plantsunder essentially non-existent pathogen pressure, wherein the plants,the plant propagules, the seed of the plants and/or the locus where theplants are growing or are intended to grow are treated with an effectiveamount of a composition comprising

-   -   a) the Bacillus subtilis strain with NRRL Accession No. B-21661        or a cell-free extract thereof, and/or a mutant of this strain        or extract having all the identifying characteristics of the        respective strain or extract as component (I), and    -   b) two chemical compounds as component (II), selected from the        active compound groups A) to J) as defined above.

In another preferred embodiment, the present invention relates to amethod for increasing the vigor and/or crop yield of agricultural plantsunder essentially non-existent pathogen pressure, wherein the plants,the plant propagules, the seed of the plants and/or the locus where theplants are growing or are intended to grow are treated with an effectiveamount of a composition comprising

-   -   a) the Bacillus subtilis strain with NRRL Accession No. B-21661        or a cell-free extract thereof, and/or a mutant of this strain        or extract having all the identifying characteristics of the        respective strain or extract as component (I) and    -   b) one chemical compound as component (II), selected from the        active compound groups A) to J) as defined in claim 1        are applied simultaneously, that is jointly or separately, or in        succession.

Component (I) embraces not only the isolated, pure cultures of theBacillus subtilis strain or a cell-free extract thereof, but also theirsuspensions in a whole broth culture or a metabolite-containingsupernatant or a purified metabolite obtained from a whole broth cultureof the strain.

The Bacillus subtilis strain, extracts and mutants thereof, and themetabolites produced by this strain, its preparation and action againstharmful fungi is known from WO 98/50422 and WO 00/29426, therein alsoreferred to as AQ 713 (QST 713). Said strain, however, may also bereferred to in the prior art as Bacillus amyloliquefaciens.

SERENADE® is a microbial biological control agent based on Bacillussubtilis which protects against fungal and bacterial plant pathogens.Bacillus subtilis strain QST 713 is a naturally occurring widespreadbacterium that can be used to control plant diseases including blight,scab, gray mold, and several types of mildew. Regulatory authorities inUSA and Europe classified Bacillus subtilis QST 713 as displaying noadverse effects on humans or the environment. The bacterium, Bacillussubtilis, is prevalent in soils and has been found in a variety ofhabitats worldwide. The QST 713 strain of Bacillus subtilis is known tobe antagonistic towards many fungal plant pathogens.

This antagonism may be achieved in several ways including nutrientcompetition, site exclusion, colonization, and attachment of thebacteria to the fungal pathogen. In addition, the QST 713 strain ofBacillus subtilis might induce plant's natural systemic resistance orsystemic acquired resistance (SAR) against bacterial pathogens. OST 713can stop plant pathogen spores from germinating, disrupt germ tubegrowth, and inhibit attachment of the plant pathogen to the leaf.

Suitable formulations of the Bacillus subtilis strain with NRRLAccession No. B-21661 are commercially available under the tradenamesSERENADE®, SERENADE® MAX and SERENADE® ASO from AgraQuest, Inc., 1540Drew Avenue, Davis, Calif. 95618, U.S.A.

In one embodiment, a commercially available formulation of the aboveidentified Bacillus subtilis strain is used.

A reduction of yield loss due to the control of fungal pathogens bycomponent (I) is well known (see, for example Highland (2002): Proc.Fla. State Hort. Soc.: 115, 186-188).

In a presentation at the 2008 Acorbat conference, results were presentedshowing that when Bacillus subtilis QST 713, formulated as the SERENADE®product, was applied to black sigatoka-infected bananas, it controlleddisease comparable to the chemical standard (mancozeb) resulting in anincreased banana bunch production by 30% above the mancozeb-treatedplots (see Manker and Seiler (2008): “Bacillus subtilis strain QST 713as an Alternative Protectant Multi-Site Fungicide for SustainableControl of Black Sigatoka in Banana Production” distributed Nov. 10,2008 at Acorbat conference in Guayaquil, Ecuador).

However, the fact that the application of the Bacillus subtilis strainwith NRRL Accession No. B-21661 or a cell-free extract thereof, and/or amutant of this strain or extract having all the identifyingcharacteristics of the respective strain or extract increases the vigorand/or the yield of agricultural plants even under essentiallynon-existant pathogen pressure is new and surprising because it couldnot have been expected that the yield and/or vigor increase wouldgenerally be above the level that could be reached by combating thephytopathogenic fungi and/or bacteria known to reduce a crop's vigor andyield. This is especially true with respect to the application ofcompositions comprising component (I) and at least one component (II)which have shown to be able to synergistically increase of a crop'svigor and yield according to the invention.

The term “essentially non-existant pathogen pressure” refers to asituation in which pathogens are present within the area of growth of aplant but in a quantity that is not harmful to the plant and which doesneither result in a decrease of vigor nor in a decrease of yield.

In view of the increasing world population of humans, it becomes moreand more important to increase the worldwide food production (yield) andfood quality which is based upon healty plants that display high levelsof vigor.

Accordingly, it was an object of the present invention to provide agentswhich increase the vigor and/or yield of plants to an extent which ismore than healthy plants under essentially non-existent pathogenpressure would produce, whereas the term pathogen shall primarely meanfungal pathogens and optionally bacterial pathogens causing damage toplants, preferably both fungal and bacterial pathogens.

We have found that this object is achieved by applying component (I)and, optionally at least one component (II) as defined at the outset. Ina preferred embodiment of the invention, component (I) and at least onecomponent (II) as defined at the outset, are applied. By simultaneous,that is joint or separate, application of component (I) and at least onecomponent (II), the vigor and/or the yield of agricultural plants may beincreased in a superadditive that means synergistic manner.

The concept of using biopesticides such as a SERENADE® product incombination with chemicals is new and has a number of benefits. One ofthe most important is the fact that biopesticides such as SERENADE®don't leave any chemical residues on the crops, meaning that they can beused right up to the day of harvest. Crops are usually left perilouslyunprotected in the days leading up to harvest, as conventionalpesticides cannot be applied during this time. Accordingly, in apreferred embodiment of the method according to the invention, at leastone component (II) selected from the active compound groups (A) to (J)are applied before the Pre-Harvest Intervall while compond (I) isapplied during the Pre-Harvest Interval!.

The term “Pre-Harvest Interval” is to be understood as the time betweenthe last pesticide application (component II) and harvest of the treatedcrops.

The term “principal growth stage” refers to the extended BBCH-scalewhich is a system for a uniform coding of phenologically similar growthstages of all mono- and dicotyledonous plant species in which the entiredevelopmental cycle of the plants is subdivided into clearlyrecognizable and distinguishable longer-lasting developmental phases.The BBCH-scale uses a decimal code system, which is divided intoprincipal and secondary growth stages. The abbreviation BBCH derivesfrom the Federal Biological Research Centre for Agriculture and Forestry(Germany), the Bundessortenamt (Germany) and the chemical industry. ABBCH value of 23/29 indicates that the plants measured had reached agrowing stadium in between 23 and 29.

NRRL is the abbreviation for the Agricultural Research Service CultureCollection, an international depositary authority for the purposes ofdeposing microorganism strains under the BUDAPEST TREATY ON THEINTERNATIONAL RECOGNITION OF THE DEPOSIT OF MICROORGANISMS FOR THEPURPOSES OF PATENT PROCEDURE, having the address National Center forAgricultural Utilization Research, Agricultural Research Service, U.S.Department of Agriculture, 1815 North University Street, Peoria, Ill.61604, USA.

Component (I) embraces not only the isolated, pure cultures of theBacillus subtilis strain, but also its suspensions in a whole brothculture or as a metabolite-containing supernatant or a purifiedmetabolite obtained from a whole broth culture of the strain.

“Whole broth culture” refers to a liquid culture containing both cellsand media.

“Supernatant” refers to the liquid broth remaining when cells grown inbroth are removed by centrifugation, filtration, sedimentation, or othermeans well known in the art.

The term “metabolite” refers to any compound, substance or byproductproduced by a microorganism (such as fungi and bacteria) that hasfungicidal activity.

The chemical compounds mentioned above as component (II) are generallyknown (cf., for example, http://www.hclrss.demon.co.uk/index.html); mostof them are commercially available. Their pesticidal action and methodsfor producing them are also known. For instance, the commerciallyavailable compounds may be found in The Pesticide Manual, 14th Edition,British Crop Protection Council (2006) among other publications.

Bixafen is known from WO 03/070705; penflufen is known from WO03/010149;Sedaxane is known from WO 03/074491; they can be prepared inthe manner described therein. Isopyrazam is known from WO 04/035589 andcan be prepared in the manner described therein or as described in WO2007/068417.N-(3′,4′,5′-trifluorobiphenyl-2-yl)-3-difluoromethyl-1-methyl-1H-pyrazole-4-carboxamideis known from WO 06/087343 and can be prepared in the manner describedtherein. Metrafenone,3′-bromo-2,3,4,6′-tetramethoxy-2′,6-dimethylbenzophenone, is known fromU.S. Pat. No. 5,945,567.

The compounds according to the invention can be present in differentcrystal modifications whose biological activity may differ. They arelikewise subject matter of the present invention.

Preference is given to the application of component (I) in combinationwith at least one component (II).

Accordingly, in a preferred embodiment of the method according to theinvention, component (I) is applied together with a component (II)selected from the groups A), B), C), D), E), F) and G). In a morepreferred embodiment of the method according to the invention, component(I) is applied together with a component (II) selected from the groupsA), B), C), E) and G). In an even more preferred embodiment of themethod according to the invention, component (I) is applied togetherwith a component (II) selected from the groups A) and B). In a mostpreferred embodiment of the method according to the invention, component(I) is applied together with a component (II) selected from the groupA).

In one embodiment of the method according to the invention, component(I) is applied together with a component (II) selected from group A)(strobilurins) consisting of azoxystrobin, dimoxystrobin, enestroburin,fluoxastrobin, kresoxim-methyl, metominostrobin, orysastrobin,picoxystrobin, pyraclostrobin, pyribencarb, trifloxystrobin,2-(2-(6-(3-chloro-2-methyl-phenoxy)-5-fluoro-pyrimidin-4-yloxy)-phenyl)-2-methoxy

imino-N-methyl-acetamide,3-methoxy-2-(2-(N-(4-methoxy-phenyl)-cyclopropane-carboximidoyl

sulfanylmethyl)-phenyl)-acrylic acid methyl ester,methyl(2-chloro-5[1-(3-methylbenzyloxyimino)

ethyl]benzyl)

carbamate and 2(2-(3-(2,6-di

chlorophenyl)-1-methyl-allylidene

aminooxy

methyl)-phenyl)-2-methoxyimino-N methyl-acetamide. Among the group A)comprising strobilurins as component (II), azoxystrobin, dimoxystrobin,enestroburin, fluoxastrobin, kresoxim-methyl, orysastrobin,picoxystrobin, pyraclostrobin and trifloxystrobin are preferred.Azoxystrobin, dimoxystrobin, and pyraclostrobin are especiallypreferred. Pyraclostrobin is most preferred.

In a especially preferred embodiment of the invention, component (I) isapplied with pyraclostrobin as component (II).

In another especially preferred embodiment of the invention, component(I) is applied with epoxiconazole and pyraclostrobin as component (II).

In another embodiment of the method according to the invention,component (I) is applied together with a component (II) selected fromgroup B) (carboxamides) consisting of

-   -   carboxanilides selected from benalaxyl, benalaxyl-M, benodanil,        bixafen, boscalid, carboxin, fenfuram, fenhexamid, flutolanil,        furametpyr, isopyrazam, isotianil, kiralaxyl, mepronil,        metalaxyl, metalaxyl-M (mefenoxam), ofurace, oxadixyl,        oxycarboxin, penthiopyrad, sedaxane, tecloftalam, thifluzamide,        tiadinil, 2-amino-4-methyl-thiazole-5-carboxanilide, 2        chloro-N(1,1,3-trimethylindan-4-yl)-nicotinamide, N-(3′,4′,5′        trifluorobiphenyl-2 yl)-3-difluoromethyl-1-methyl-1H-pyrazole-4        carboxamide, N-(4′-trifluoromethylthiobiphenyl-2-yl)-3        difluoromethyl-1-methyl-1H-pyrazole-4-carboxamide, N(2-(1,3        dimethyl-butyl)-phenyl)-1,3-dimethyl-5-fluoro-1H-pyrazole-4-carboxamide        and        N-(2-(1,3,3-trimethyl-butyl)-phenyl)-1,3-dimethyl-5-fluoro-1H-pyrazole-4        carboxamide;    -   carboxylic morpholides selected from dimethomorph, flumorph and        pyrimorph;    -   benzoic acid amides selected from flumetover, fluopicolide,        fluopyram, zoxamide and        N-(3-Ethyl-3,5,5-trimethyl-cyclohexyl)-3-formylamino-2-hydroxy-benzamide;    -   other carboxamides selected from carpropamid, dicyclomet,        mandiproamid, oxytetracyclin, silthiofarm and        N-(6-methoxy-pyridin-3-yl)cyclopropanecarboxylic acid amide.

Among the group B) comprising carboxamides as component (II),carboxanilides, carboxylic morpholides and benzoic acid amides arepreferred. Within the group of carboxanilides, bixafen, boscalid andN-(3′,4′,5′-trifluorobiphenyl-2-yl)-3-difluoromethyl-1-methyl-1H-pyrazole-4-carboxamideare especially preferred. Within the group of carboxylic morpholides,dimethomorph and flumorph are especially preferred. Within the group ofbenzoic acid amides, zoxamide is especially preferred. Bixafen, boscalidandN-(3′,4′,5′-trifluorobiphenyl-2-yl)-3-difluoromethyl-1-methyl-1H-pyrazole-4-carboxamideare even more preferred.N-(3′,4′,5′-trifluorobiphenyl-2-yl)-3-difluoromethyl-1-methyl-1H-pyrazole-4-carboxamideis most preferred.

In a especially preferred embodiment of the invention, component (I) isapplied with boscalid as component (II). In another especially preferredembodiment of the invention, component (I) is applied withN-(3′,4′,5′-trifluorobiphenyl-2-yl)-3-difluoromethyl-1-methyl-1H-pyrazole-4-carboxamideas component (II).

In another embodiment of the method according to the invention,component (I) is applied together with a component (II) selected fromgroup C) (azoles) consisting of

-   -   triazoles selected from azaconazole, bitertanol, bromuconazole,        cyproconazole, difenoconazole, diniconazole, diniconazole-M,        epoxiconazole, fen-buconazole, fluquinconazole, flusilazole,        flutriafol, hexaconazole, imibenconazole, ipconazole,        metconazole, myclobutanil, oxpocona-zole, paclobutrazole,        penconazole, propiconazole, prothioconazole, simeconazole,        tebuconazole, tetraconazole, triadimefon, triadimenol,        triticonazole, uniconazole and        1-(4-chloro-phenyl)-2-([1,2,4]triazol-1-yl)-cycloheptanol;    -   imidazoles selected from cyazofamid, imazalil, pefurazoate,        prochloraz and triflumizol;    -   benzimidazoles selected from benomyl, carbendazim, fuberidazole        and thiabendazole;    -   others selected from ethaboxam, etridiazole, hymexazole and        2-(4-chlorophenyl)-N-[4-(3,4-dimethoxy-phenyl)-isoxazol-5-yl]-2-prop-2-ynyloxy-acetamide.

Among group C) comprising azoles as component (II), triazoles,imidazoles, benzimidazoles and ethaboxam are preferred. Within the groupof triazoles, bitertanol, cyproconazole, difenoconazole, epoxiconazole,fenbuconazole, fluquinconazole, flusilazole, flutriafol, hexaconazole,metconazole, myclobutanil, propiconazole, tebuconazole and triticonazoleare especially preferred. Within the group of imidazole, cyazofamid andprochloraz are especially preferred. Within the group of benzimidaoles,benomyl, carbendazim and thiabendazole are especially preferred. Withingroup C), cyproconazole, difenoconazole, epoxiconazole and tebuconazoleare especially preferred. Epoxiconazole is most preferred.

In an especially preferred embodiment of the invention, component (I) isapplied with epoxiconazole as component (II).

In another especially preferred embodiment of the invention, component(I) is applied with difenoconazole and mefenoxam as component (II).

In another embodiment of the method according to the invention,component (I) is applied together with a component (II) selected fromthe group D) (heterocyclic compounds) consisting of

-   -   pyridines selected from fluazinam, pyrifenox,        3-[5-(4-chloro-phenyl)-2,3-dimethyl-isoxazolidin-3 yl]-pyridine,        3-[5-(4-methyl-phenyl)-2,3-dimethyl-isoxazolidin-3-yl]-pyridine,        2,3,5,6-tetra-chloro-4-methanesulfonyl-pyridine,        3,4,5-trichloropyridine-2,6-di-carbonitrile,        N-(1-(5-bromo-3-chloro-pyridin-2-yl)-ethyl)-2,4-dichloronicotinamide        and N[(5        bromo-3-chloro-pyridin-2-yl)-methyl]-2,4-dichloro-nicotinamide;    -   pyrimidines selected from bupirimate, cyprodinil, diflumetorim,        fenarimol, ferimzone, mepanipyrim, nitrapyrin, nuarimol,        pyrimethanil;    -   piperazines: triforine;    -   pyrroles selected from fenpiclonil and fludioxonil;    -   morpholines selected from aldimorph, dodemorph,        dodemorph-acetate, fen-propimorph and tridemorph;    -   piperidines: fenpropidin;    -   dicarboximides selected from fluoroimid, iprodione, procymidone        and vinclozolin;    -   non-aromatic 5-membered heterocycles selected from famoxadone,        fenamidone, flutianil, octhilinone, probenazole and        5-amino-2-isopropyl-3-oxo-4-orthotolyl-2,3-dihydro-pyrazole-1        carbothioic acid S-allyl ester;    -   others selected from acibenzolar-S-methyl, amisulbrom, anilazin,        blasticidin-S, captafol, captan, chinomethionat, dazomet,        debacarb, diclomezine, difenzoquat, difenzoquat-methyl        sulfate, fenoxanil, Folpet, oxolinic acid, piperalin,        proquinazid, pyroquilon, quin        oxyfen, triazoxide, tricy-clazole,        2-butoxy-6-iodo-3-propylchromen-4-one, 5-chloro-1        (4,6-dimethoxy-pyrimidin-2-yl)-2-methyl-1H-benzoimidazole, 5        chloro-7 (4-methyl        piperidin-1-yl)-6-(2,4,6-trifluorophenyl)-[1,2,4]triazolo        [1,5 a]pyrimidine and 5-ethyl-6 octyl[1,2,4]triazolo[1,5-a]pyri        midine-7 ylamine.

Among group D) comprising heterocyclic compounds as component (II),pyridines, pyrimidines, morpholines, piperidines, dicarboximides andnon-aromatic 5-membered heterocycles are preferred. Within the group ofpyridines, fluazinam is especially preferred. Within the group ofpyrimidines, cyprodinil, fenarimol and pyrimethanil are especiallypreffered. Within the group of morpholines, dodemorph-acetate,fenpropimorph and tridemorph are especially preferred. Within the groupof piperidines, fenpropidin is especially preferred. Within the group ofdicarboximides, iprodione is especially preferred. Within the group ofnon-aromatic 5-membered heterocycles famoxadone and fenamidone areespecially preferred. In addition, samisulbrom, Folpet, proquinazid andquinoxyfen are especially preferred. Cyprodinil, fenpropidin, iprodione,famoxadone, fenamidone, amisulbrom, proquinazid, quinoxyfen and Folpetare even more preferred. Fenpropimorph, tridemorph and fenpropidin aremost peferred.

In another embodiment of the method according to the invention,component (I) is applied together with a component (II) selected fromgroup E) (carbamates) consisting of

-   -   thio- and dithiocarbamates selected from ferbam, mancozeb,        maneb, metam, methasulphocarb, metiram, propineb, thiram, zineb        and ziram;    -   carbamates selected from benthiavalicarb, diethofencarb,        iprovalicarb, propamo-carb, propamocarb hydrochlorid, valiphenal        and N-(1-(1-(4-cyano-phenyl)        ethanesulfonyl)-but-2-yl)carbamic acid-(4-fluorophenyl)ester.

Among group E) comprising carbamates as component (II), thio- anddithiocarbamates and carbamates are preferred. Within the group of thio-and dithiocarbamates, mancozeb, maneb, metiram, propineb, thiram, zineband ziram are more preferred. Within the group of carbamates,benthiavalicarb, iprovalicarb, valiphenal and propamocarb and valiphenalare preferred. Mancozeb, metiram and propineb are even more preferred.

In another embodiment of the method according to the invention,component (I) is applied together with a component (II) selected fromthe group F). Among the group F) comprising other active substances ascomponent (II), antibiotics, sulfur-containing heterocyclyl compounds,inorganic active substances, cymoxanil, metrafenone, spiroxamine fentinacetate, fentin chloride and fentin hydroxide are preferred. Within thegroup of antibiotics, kasugamycin, kasugamycin hydrochloride-hydrate andstreptomycin are especially preferred. Within the group ofsulfur-containing heterocyclyl compounds, dithianon is especiallypreferred. In addition, cymoxanil, metrafenone, spiroxamine, fentinacetate, fentin chloride, fentin hydroxide are preferred. Inorganicactive substances selected from Bordeaux mixture, copper acetate, copperhydroxide, copper oxychloride, basic copper sulfate and sulfur areespecially preferred.

In an especially preferred embodiment of the invention, component (I) isapplied together with component (II) selected from Bordeaux mixture,copper acetate, copper hydroxide, copper oxychloride, basic coppersulfate and sulfur.

In another embodiment of the method according to the invention,component (I) is applied together with a component (II) selected fromgroup G) (plant growth regulators; PGRs) consisting of abscisic acid,amidochlor, ancymidol, 6-benzylaminopurine, brassinolide, butralin,chlormequat (chlormequat chloride), choline chloride, cyclanilide,daminozide, dikegulac, dimethipin, 2,6-dimethylpuridine, ethephon,flumetralin, flurprimidol, fluthiacet, forchlorfenuron, gibberellicacid, inabenfide, indole-3-acetic acid, maleic hydrazide, mefluidide,mepiquat (mepiquat chloride), naph-thaleneacetic acid, N 6benzyladenine, paclobutrazol, prohexadione (prohexadione-calcium),prohydrojasmon, thidiazuron, triapenthenol, tributylphosphorotrithioate, 2,3,5 tri iodobenzoic acid, trinexapac-ethyl anduniconazole. Among the group G) comprising plant growth regulators(PGRs) as component (II), chlormequat (chlormequat chloride), mepiquat(mepiquat chloride and prohexadione (prohexadione-calcium) arepreferred.

One indicator for the condition of the plant is its yield. “Yield” is tobe understood as any plant product of economic value that is produced bythe plant such as grains, fruits in the proper sense, vegetables, nuts,grains, seeds, wood (e.g. in the case of silviculture plants) or evenflowers (e.g. in the case of gardening plants, ornamentals). The plantproducts may in addition be further utilized and/or processed afterharvesting.

According to the present invention, “increased yield” of a plant, inparticular of an agricultural, silvicultural and/or ornamental plantmeans that the yield of a product of the respective plant is increasedby a measurable amount over the yield of the same product of the plantproduced under the same conditions, but without the application of thecomposition of the invention. Increased yield can be characterized,among others, by following improved properties of the plant:

-   -   increased plant weight,    -   increased plant height,    -   increased biomass such as higher fresh and/or dry weight    -   higher grain yield    -   more tillers    -   larger leaves    -   increased shoot growth    -   increased protein content    -   increased oil content    -   increased starch content    -   increased pigment content

According to one embodiment of the present invention, the yield isincreased by at least 5%. According to another embodiment of the presentinvention, the yield is increased by least 10%. According to anotherembodiment of the present invention, the yield is increased by least15%. According to another embodiment of the present invention, the yieldis increased by least 30%. According to another embodiment of thepresent invention, the yield is increased by least 40%.

Another indicator for the condition of the plant is the “plant vigor”.The plant vigor becomes manifest in several aspects such as the generalvisual appearance. Improved plant vigor can be characterized, amongothers, by following improved properties of the plant:

-   -   improved vitality of the plant,    -   improved plant growth,    -   improved plant development,    -   improved visual appearance,    -   improved plant stand (less plant verse/lodging),    -   improved emergence,    -   enhanced root growth and/or more developed root system,    -   enhanced nodulation, in particular rhizobial nodulation,    -   bigger leaf blade,    -   increased plant size,    -   increased plant weight,    -   increased plant height,    -   increased tiller number,    -   increased shoot growth,    -   increased root growth (extensive root system),    -   increased size of root mass (extensive root system),    -   increased yield when grown on poor soils or unfavorable climate,    -   enhanced photosynthetic activity    -   change of color (e.g. enhanced pigment content (e.g. Chlorophyll        content)    -   earlier flowering,    -   earlier fruiting,    -   earlier and improved germination,    -   earlier (advanced) grain maturity,    -   improved self-defence mechanisms    -   less non-productive tillers,    -   less dead basal leaves,    -   less input needed (such as fertilizers or water)    -   greener leaves and increased green leaf area    -   complete maturation under shortened vegetation periods    -   less fertilizers needed,    -   less seeds needed,    -   easier harvesting    -   faster and more uniform ripening    -   longer shelf-life    -   longer panicles,    -   delay of senescence,    -   stronger and/or more productive tillers,    -   better extractability of ingredients    -   improved quality of seeds (for being seeded in the following        seasons for seed production)    -   reduced production of ethylene and/or the inhibition of its        reception by the plant    -   spindliness of leaves    -   inceased number of ears/m²

The improvement of the plant vigor according to the present inventionparticularly means that the improvement of any one or several or all ofthe above mentioned plant characteristics are improved independently ofthe pesticidal action of the composition or active ingredients. Anincreased vigor may for example result in a higher percentatage ofplants that can be transplanted to the field or an increased number ofmarketable plants (such as tomatoes).

The term “plants” is to be understood as plants of economic importanceand/or men-grown plants such as cultivated plants. They are preferablyselected from agricultural, silvicultural and horticultural (includingornamental) plants. The term “plant” as used herein includes all partsof a plant such as germinating seeds, emerging seedlings, herbaceousvegetation as well as established woody plants including all belowgroundportions (such as the roots) and aboveground portions.

Generally the term “plants” also includes plants which have beenmodified by breeding, mutagenesis or genetic engineering. Geneticallymodified plants are plants, which genetic material has been modified bythe use of recombinant DNA techniques. The use of recombinant DNAtechniques makes modifications possible that cannot readily be obtainedby cross breeding under natural circumstances, mutations or naturalrecombination.

Agricultural plants which may exhibit an increase in vigor and/or cropyield are, for example, cereals, for example wheat, rye, barley,triticale, oats or rice; beet, for example sugar beet or fodder beet;fruits, such as pomes, stone fruits or soft fruits, for example apples,pears, plums, peaches, almonds, cherries, strawberries, raspberries,blackberries or gooseberries; leguminous plants, such as lentils, peas,alfalfa or soybeans; oil plants, such as rape, mustard, olives,sunflowers, coconut, cocoa beans, castor oil plants, oil palms, groundnuts or soybeans; cucurbits, such as squashes, cucumber or melons; fiberplants, such as cotton, flax, hemp or jute; citrus fruit, such asoranges, lemons, grapefruits or mandarins; vegetables, such as broccoli,spinach, lettuce, asparagus, cabbages, carrots, onions, tomatoes,potatoes, cucurbits or paprika; lauraceous plants, such as avocados,cinnamon or camphor; energy and raw material plants, such as corn,soybean, rape, sugar cane or oil palm; corn; tobacco; nuts; coffee; tea;bananas; vines (table grapes and grape juice grape vines); hop; turf;natural rubber plants or ornamental and forestry plants, such asflowers, shrubs, broad-leaved trees or evergreens, for example conifers;and on the plant propagation material, such as seeds, and the cropmaterial of these plants.

Agricultural plants which exhibit vigor and/or crop yield increase arein particular bananas, broccoli, tomatoes, pepper and wheat.

In a preferred embodiment of the invention, the yield and/or vigor isincreased in an agricultural plant selected from soybean, corn, wheat,triticale, barley, oat, rye, rape, millet, rice, sunflower, cotton,sugar beet, pome fruit, stone fruit, citrus, banana, strawberry,blueberry, almond, grape, mango, papaya, peanut, potato, tomato, pepper,cucurbit, cucumber, melon, watermelon, garlic, onion, broccoli, carrot,cabbage, bean, dry bean, canola, pea, lentil, alfalfa, trefoil, clover,flax, elephant grass, grass, lettuce, sugarcane, tea, tobacco andcoffee; each in its natural or genetically modified form

In a preferred embodiment of the invention, the yield and/or vigor isincreased in grapes, fruits such as pomes, stone fruits or soft fruits,for example apples, pears, plums, peaches, almonds, cherries,strawberries, raspberries, blackberries or gooseberries and/orvegetables such as broccoli, spinach, lettuce, asparagus, cabbages,carrots, onions, tomatoes, potatoes, cucurbits or paprika.

In an especially preferred embodiment of the invention, the yield and/orvigor is increased in bananas and/or grapes.

The term “plant propagation material” is to be understood to denote allthe generative parts of the plant such as seeds and vegetative plantmaterial such as cuttings and tubers (e.g. potatoes), which can be usedfor the multiplication of the plant. This includes seeds, roots, fruits,tubers, bulbs, rhizomes, shoots, sprouts and other parts of plants.Seedlings and young plants, which are to be transplanted aftergermination or after emergence from soil, may also be mentioned. Theseyoung plants may also be treated totally or partially by immersion orpouring before transplantation.

The term “cultivated plants” is to be understood as including plantswhich have been modified by breeding, mutagenesis or geneticengineering. Genetically modified plants are plants, which geneticmaterial has been so modified by the use of recombinant DNA techniquesthat under natural circumstances cannot readily be obtained by crossbreeding, mutations or natural recombination. Typically, one or moregenes have been integrated into the genetic material of a geneticallymodified plant in order to improve certain properties of the plant.

The term “cultivated plants” is to be understood also including plantsthat have been rendered tolerant to applications of specific classes ofherbicides, such as hydroxyphenylpyruvate dioxygenase (HPPD) inhibitors;acetolactate synthase (ALS) inhibitors, such as sulfonyl ureas (see e.g.U.S. Pat. No. 6,222,100, WO 01/82685, WO 00/26390, WO 97/41218, WO98/02526, WO 98/02527, WO 04/106529, WO 05/20673, WO 03/14357, WO03/13225, WO 03/14356, WO 04/16073) or imidazolinones (see e.g. U.S.Pat. No. 6,222,100, WO 01/82685, WO 00/26390, WO 97/41218, WO 98/02526,WO 98/02527, WO 04/106529, WO 05/20673, WO 03/14357, WO 03/13225, WO03/14356, WO 04/16073); enolpyruvylshikimate-3-phosphate synthase(EPSPS) inhibitors, such as glyphosate (see e.g. WO 92/00377); glutaminesynthetase (GS) inhibitors, such as glufosinate (see e.g. EP-A-0242236,EP-A-242246) or oxynil herbicides (see e.g. U.S. Pat. No. 5,559,024) asa result of conventional methods of breeding or genetic engineering.Several cultivated plants have been rendered tolerant to herbicides byconventional methods of breeding (mutagenesis), for example Clearfield®summer rape (Canola) being tolerant to imidazolinones, e.g. imazamox.Genetic engineering methods have been used to render cultivated plants,such as soybean, cotton, corn, beets and rape, tolerant to herbicides,such as glyphosate and glufosinate, some of which are commerciallyavailable under the trade names RoundupReady® (glyphosate) andLibertyLink® (glufosinate).

The term “cultivated plants” is to be understood also including plantsthat are by the use of recombinant DNA techniques capable to synthesizeone or more insecticidal proteins, especially those known from thebacterial genus Bacillus, particularly from Bacillus thuringiensis, suchas δ-endotoxins, e.g. CryIA(b), CryIA(c), CryIF, CryIF(a2), CryIIA(b),CryIIIA, CryIIIB(b1) or Cry9c; vegetative insecticidal proteins (VIP),e.g. VIP1, VIP2, VIP3 or VIP3A; insecticidal proteins of bacteriacolonizing nematodes, for example Photorhabdus spp. or Xenorhabdus spp.;toxins produced by animals, such as scorpion toxins, arachnid toxins,wasp toxins, or other insect-specific neurotoxins; toxins produced byfungi, such Streptomycetes toxins, plant lectins, such as pea or barleylectins; agglutinins; proteinase inhibitors, such as trypsin inhibitors,serine protease inhibitors, patatin, cystatin or papain inhibitors;ribosome-inactivating proteins (RIP), such as ricin, maize-RIP, abrin,luffin, saporin or bryodin; steroid metabolism enzymes, such as3-hydroxysteroid oxidase, ecdysteroid-IDP-glycosyl-transferase,cholesterol oxidases, ecdysone inhibitors or HMG-CoA-reductase; ionchannel blockers, such as blockers of sodium or calcium channels;juvenile hormone esterase; diuretic hormone receptors (helicokininreceptors); stilben synthase, bibenzyl synthase, chitinases orglucanases. In the context of the present invention these insecticidalproteins or toxins are to be understood expressly also as pre-toxins,hybrid proteins, truncated or otherwise modified proteins. Hybridproteins are characterized by a new combination of protein domains,(see, for example WO 02/015701). Further examples of such toxins orgenetically modified plants capable of synthesizing such toxins aredisclosed, for example, in EP-A374753, WO 93/007278, WO 95/34656, EP-A427529, EP-A451878, WO 03/018810 and WO 03/052073. The methods forproducing such genetically modified plants are generally known to theperson skilled in the art and are described, for example, in thepublications mentioned above. These insecticidal proteins contained inthe genetically modified plants impart to the plants producing theseproteins tolerance to harmful pests from all taxonomic groups ofathropods, especially to beetles (Coeloptera), two-winged insects(Diptera), and moths (Lepidoptera) and to nematodes (Nematoda).

Genetically modified plants capable to synthesize one or moreinsecticidal proteins are, for example, described in the publicationsmentioned above, and some of which are commercially available such asYieldGard® (corn cultivars producing the CryIAb toxin), YieldGard® Plus(corn cultivars producing Cry1Ab and Cry3Bb1 toxins), Starlink® (corncultivars producing the Cry9c toxin), Herculex® RW (corn cultivarsproducing Cry34Ab1, Cry35Ab1 and the enzymePhosphinothricin-N-Acetyltransferase [PAT]); NuCOTN® 33B (cottoncultivars producing the Cry1Ac toxin), Bollgard® I (cotton cultivarsproducing the Cry1Ac toxin), Bollgard® II (cotton cultivars producingCry1Ac and Cry2Ab2 toxins); VIPCOT® (cotton cultivars producing aVIP-toxin); NewLeaf® (potato cultivars producing the Cry3A toxin);Bt-Xtra®, NatureGard®, KnockOut®, BiteGard®, Protecta®, Bt11 (e.g.Agrisure® CB) and Bt176 from Syngenta Seeds SAS, France, (corn cultivarsproducing the Cry1Ab toxin and PAT enyzme), MIR604 from Syngenta SeedsSAS, France (corn cultivars producing a modified version of the Cry3Atoxin, c.f. WO 03/018810), MON 863 from Monsanto Europe S.A., Belgium(corn cultivars producing the Cry3Bb1 toxin), IPC531 from MonsantoEurope S.A., Belgium (cotton cultivars producing a modified version ofthe Cry1Ac toxin) and 1507 from Pioneer Overseas Corporation, Belgium(corn cultivars producing the Cry1F toxin and PAT enzyme).

The term “cultivated plants” is to be understood also including plantsthat are by the use of recombinant DNA techniques capable to synthesizeone or more proteins to increase the resistance or tolerance of thoseplants to bacterial, viral or fungal pathogens. Examples of suchproteins are the so-called “pathogenesis-related proteins” (PR proteins,see, for example EP-A 0392225), plant disease resistance genes (forexample potato cultivars, which express resistance genes acting againstPhytophthora infestans derived from the mexican wild potato Solanumbulbocastanum) or T4-lysozym (e.g. potato cultivars capable ofsynthesizing these proteins with increased resistance against bacteriasuch as Erwinia amylvora). The methods for producing such geneticallymodified plants are generally known to the person skilled in the art andare described, for example, in the publications mentioned above.

The term “cultivated plants” is to be understood also including plantsthat are by the use of recombinant DNA techniques capable to synthesizeone or more proteins to increase the productivity (e.g. biomassproduction, grain yield, starch content, oil content or proteincontent), tolerance to drought, salinity or other growth-limitingenvironmental factors or tolerance to pests and fungal, bacterial orviral pathogens of those plants.

The term “cultivated plants” is to be understood also including plantsthat contain by the use of recombinant DNA techniques a modified amountof substances of content or new substances of content, specifically toimprove human or animal nutrition, for example oil crops that producehealth-promoting long-chain omega-3 fatty acids or unsaturated omega-9fatty acids (e.g. Nexera® rape).

The term “cultivated plants” is to be understood also including plantsthat contain by the use of recombinant DNA techniques a modified amountof substances of content or new substances of content, specifically toimprove raw material production, for example potatoes that produceincreased amounts of amylopectin (e.g. Amflora® potato).

The term “protein” as used herein is to be understood as an oligopeptideor polypeptide or molecule made up of polypeptides including expresslyalso pre-proteins, hybrid proteins, peptides, truncated or otherwisemodified proteins including those derived from post-transcriptionalmodifications such as acylation (e.g. acetylation, the addition of anacetyl group, usually at the N-terminus of the protein), alkylation, theaddition of an alkyl group (e.g. addition of ethyl or methyl, usually atlysine or arginine residues) or demethylation, amidation at C-terminus,biotinylation (acylation of conserved lysine residues with a biotinappendage), formylation, γ-carboxylation dependent on Vitamin K,glutamylation (covalent linkage of glutamic acid residues),glycosylation (addition of a glycosyl group to either asparagine,hydroxylysine, serine, or threonine, resulting in a glycoprotein),glycation (nonenzymatic attachment of sugars), glycylation (covalentlinkage of one to more glycine residues), covalent attachment of a hememoiety, hydroxylation, iodination, isoprenylation (addition of anisoprenoid group such as farnesol and geranylgeraniol), lipoylation(attachment of a lipoate functionality) including prenylation, GPIanchor formation (e.g. myristoylation, farnesylation andgeranylgeranylation), covalent attachment of nucleotides or derivativesthereof including ADP-ribosylation and flavin attachment, oxidation,pegylation, covalent attachment of phosphatidylinositol,phosphopantetheinylation (addition of a 4′-phosphopantetheinyl moietyfrom coenzyme A), phosphorylation (addition of a phosphate group,usually to serine, tyrosine, threonine or histidine), pyroglutamateformation, racemization of proline, tRNA-mediated addition of aminoacids such as arginylation, sulfation (addition of a sulfate group to atyrosine), selenoylation (co-translational incorporation of selenium inselenoproteins), ISGylation (covalent linkage to the ISG15 protein[Interferon-stimulated Gene 15]), SUMOylation (covalent linkage to theSUMO protein [Small Ubiquitin-related MOdifier]), ubiquitination(covalent linkage to the protein ubiquitin or poly-ubiquitin),citrullination or deimination (conversion of arginine to citrulline),deamidation (conversion of glutamine to glutamic acid or asparagine toaspartic acid), formation of disulfide bridges (covalent linkage of twocysteine amino acids) or proteolytic cleavage (cleavage of a protein ata peptide bond).

The term “locus” is to be understood as any type of environment, soil,area or material where the plant is growing or intended to grow as wellas the environmental conditions (such as temperature, wateravailability, radiation) that have an influence on the growth anddevelopment of the plant and/or its propagules. In addition, the term“locus” is to be understood as a plant, seed, soil, area, material orenvironment in which a pest is growing or may grow.

“Crop yield ” is an indicator for the condition of the plant, whereas“crop” is to be understood as any plant or plant product which isfurther utilized after harvesting, e.g. fruits in the proper sense,vegetables, nuts, grains, seeds, wood (e.g. in the case of silvicultureplants), flowers (e.g. in the case of gardening plants, ornamentals)etc., that is anything of economic value that is produced by the plant.

According to the present invention, “increased yield” of a plant, inparticular of an agricultural, silvicultural and/or ornamental plantmeans that the yield of a product of the respective plant is increasedby a measurable amount over the yield of the same product of the plantproduced under the same conditions, but without the application of thecomposition of the invention.

The term “seed” embraces seeds and plant propagules of all kindsincluding but not limited to true seeds, seed pieces, suckers, corms,bulbs, fruit, tubers, grains, cuttings, cut shoots and the like andmeans in a preferred embodiment true seeds.

The term “seed treatment” comprises all suitable seed treatmenttechniques known in the art, such as seed dressing, seed coating, seeddusting, seed soaking, seed impregnation and seed pelleting.

The term “plant propagation material” or “plant propagation product” isto be understood to denote all the generative parts of the plant such asseeds and vegetative plant material such as cuttings and tubers (e.g.potatoes), which can be used for the multiplication of the plant. Thisincludes seeds, grains, roots, fruits, tubers, bulbs, rhizomes,cuttings, spores, offshoots, shoots, sprouts and other parts of plants,including seedlings and young plants, which are to be transplanted aftergermination or after emergence from soil, meristem tissues, single andmultiple plant cells and any other plant tissue from which a completeplant can be obtained.

The term “coated with” and/or “containing” generally signifies that theactive ingredient is for the most part on the surface of the propagationproduct at the time of application, although a greater or lesser part ofthe ingredient may penetrate into the propagation product, depending onthe method of application. When the said propagation product is(re)planted, it may absorb the active ingredient.

A “simultaneous” application is to be understood as the joint orseparate application of components (I) and (II).

Component (I) may be formulated with a particulate carrier substance.Said carrier may be substantially composed of water-soluble orwater-insoluble material or mixtures thereof. The Bacillus subtiliscells may be embedded into the carrier material and/or may be adsorbedto the surface of the carrier material.

The carrier substance, which may added as coformulant prior to thedrying to a suspension of usually freshly grown of the Bacillus subtiliscells, may be selected from mono-, oligo- and polysaccharides, polyols,polyethers, polymers, such as CMC or PVP, oligo- and polypeptides, fromnatural sources, such as milk, meat or cereals, derived substances ormixed substances, such as sweet whey powder, wheat semolina bran,peptone, alginates, mineral compounds, or mixtures of such materials.Said material may be dissolved in said suspension of Bacillus subtiliscells, which mxture may then be dried in order to obtain particulatematerial.

In another embodiment, said carrier may comprise a water insoluble,water-absorbent carrier substance, which may be is selected from anyorganic or inorganic material capable of removing moisture gently fromthe suspension of viable Bacillus subtilis, and in particular from thegroup consisting of zeolite, porous beads or powders, silica, groundagricultural products (as for example corn cobs), porous wood products,cellulose, cyclodextrins, and combinations thereof. The carrier may beadmixed with a suspension of usually freshly grown of Bacillus subtiliscells in order to form particulate material, which optionally my befurther subjected to drying.

In addition, additives having a stabilizing action on the Bacillussubtilis can be added to the mixture, preferably prior to thepreparation of the particulate formulation, as for example antioxidants,such as alpha-tocopherol or ascorbic acid, or mixtures thereof.Furthermore, a stabilizing action can be exerted by other substances,which are selected from inorganic salts, such as alkali metal chloridesor alkaline earth metal chlorides, inorganic or organic buffers, such asalkali metal phosphate buffer, amino acids, such as aspartic acid orglutamic acid and the salts thereof, organic carboxylic acids, such ascitric acid, organic nonvolatile solvents, such as dimethylsulfoxide,and other compounds, such as β-carotene and mixtures of these.

In a specific embodiment the particulate formulation comprises saidcarrier substance, as for example said water insoluble, water-absorbentcarrier substance, wherein said carrier substance is present in anamount of at least about 40%, as for example at lease 50, 60, 70, 80,85, 90, 95, 96, 97, 98 or 99% by the total weight of the formulation andthe Bacillus subtilis mixed with said carrier.

According to a further embodiment said particulate formulation is coatedin a manner known per se with a suitable compatible coating orencapsulating material.

Suitable encapsulating materials include, without limitation, native ormodified chitosans, native of modified starches, glucans or dextrins,celluloses modified so they are soluble, and any of a number of nativeor modified vegetable or microbial gums, including agars, guar, locust,carrageenan, xanthans, pectins, and the like, and combinations thereof.

Further suitable coating materials are polymers such as, for example,PVP, in particular a PVP product, which is commercially available underthe trade name Kollidon VA64. Another usable coating system comprises amixture of shellac and Kollidon 25 or 30, which may be supplemented withtitanium dioxide and tallow.

According to the invention, components (I) and (II) are usually employedin a weight ratio of from 1000:1 to 1:1000, as for example 200:1 to1:200, 100:1 to 1:100, as for example 90:1 to 1:90, 80:1 to 1:80, 75:1to 1:75, 50:1 to 1:50, 25:1 to 1:25 or 10:1 to 1:10.

It is preferred to employ the commercially available formulations ofcomponents (I) and (II), to which further compounds active againstharmful fungi or other pests, such as insects, arachnids or nematodes,or else herbicidal or growth-regulating active compounds (e.g. PGRs),fertilizers or sun protectants like iron oxide may be added. In apreferred embodiment, the composition comprises component (I), at leastone component (II) and iron oxide.

The further active components (II) are, if desired, added in a ratio offrom 20:1 to 1:20 to component (I).

Usually, compositions comprising component (I) and (II), whereincomponent (II) consists of only one chemical compound, are employed.However, in certain cases compositions wherein component (II) consistsof two or, if appropriate, more chemical compounds may be advantageousas well.

According to the terms of the present invention, “effective amount” isto be understood to denote all application rates for component (I) andoptionally component (II) as well as all application rates with regardto any type of mixture or composition comprising component (I) and atleast one component (II), which result in an increased vigor and/or cropyield of agricultural plants under essentially non-existent pathogenpressure. The optimal “effective amount” depends on various parameterssuch as the time of application, growth stage, area of application,application form, treated plant, soil, weather conditions etc. and mustbe determined by the person skilled in the art withing the ranges given.

In one embodiment of the method according to the invention, mixturescomprising component (I) and at least one component (II) are applied inan effective amount, whereas “effective amount” is to be understood asan amount suitable for increasing the vigor and/or crop yield ofagricultural plants under essentially non-existent pathogen pressure ina synergistic manner.

Depending on the particular components and the plants to be treated, theapplication rates for component (I) in liquid formulations are generallyfrom 0.01 l to 100 l of a composition containing the Bacillus subtilisstrain or a mutant having all the identifying characteristics of suchstrain, a cell-free extract of the strain or its mutants, or isolatedmetabolite(s) of the strain or its mutant per hectare, preferably from0.02 l to 50 l/ha, in particular from 0.05 to 18 l/ha. Application ratesfor component (I) in dry formulations are generally from 0.01 lbs/acreto 100 lbs/acre, preferably from 0.02 lbs to 50 lbs per acre and inparticular from 0.05 lbs to 5 lbs/acre. In cases in which component (I)is derived from a whole broth of the Bacillus subtilis strain or itsmutants the number of colony forming units (CFU) applied is importantand is generally from 1×1010 through lx 1015 per acre, preferably from1×1011 through 1×1014 per acre or, in particular from 1×1012 through1×1013 per acre.

Correspondingly, the application rates for component (II) are generallyfrom 1 to 2000 g/ha, 5 to 100 g/ha, preferably from 10 to 500 g/ha, inparticular from 40 to 250 g/ha of active ingredient each.

Correspondingly, the application rates for component (II) are generallyfrom 1 to 2000 g/ha, preferably from 10 to 1500 g/ha, in particular from40 to 1000 g/ha.

In a preferred embodiment of the method according to the invention, seedis treated.

The method according to the present invention is carried out by theapplication of a component (I) and optionally a component (II), or acomposition comprising components (I) and optionally a component (II),by spraying or dusting the seeds, the plants or the soils before orafter sowing of the plants or before or after emergence of the plants.

In a preferred method according to the present invention the applicationis carried out as in-furrow and/or foliar treatment. Most preferrably,the application is carried out as foliar treatment.

If an agricultural mixture according to the present invention is used inthis inventive method, the plants, the plant propagules, the seed of theplants and/or the locus where the plants are growin or are intended togrow are preferably treated simultaneously (together or separately) orsubsequently with a compenent (I) and at least one compent (II) selectedfrom the active compound groups (A) to (J).

The subsequent application is carried out with a time interval whichallows a combined action of the applied compounds. Preferably, the timeinterval for a subsequent application of component (I) and at least onecomponent (II), ranges from a few seconds up to 3 months, preferably,from a few seconds up to 1 month, more preferably from a few seconds upto 2 weeks, even more preferably from a few seconds up to 3 days and inparticular from 1 second up to 24 hours.

In a preferred embodiment, component (II) is applied before thePre-harvest interval while component (I) is applied during thePre-harvest interval.

Herein, we have found that simultaneous, that is joint or separate,application of component (I), or mixtures comprising component (I) andat least one compound selected from the active compound groups (A) to(J) or the successive application of mixtures comprising component (I),or mixtures comprising component (I) and at least one compound selectedfrom the active compound groups (A) to (J) allows increasing the vigorand/or crop yield of agricultural plants under essentially non-existentpathogen pressure compared to the control rates that are possible withthe individual compounds (synergistic mixtures).

In another preferred embodiment of the invention, component (I) or anagrochemical mixture comprising component (I) and at least one compoundselected from the active compound groups (A) to (J) is repeatedlyapplied. In one embodiment, the application is repeated two to tentimes, preferably, two to five times; most preferably three times.

The compositions according to the invention, or the single componentsseparately, can be converted into customary formulations, for examplesolutions, emulsions, suspensions, dusts, powders, pastes and granules.The use form depends on the particular intended purpose; in each case,it should ensure a fine and even distribution of the mixture accordingto the invention.

The formulations are prepared in a known manner, for example byextending the single components with solvents and/or carriers, ifdesired using emulsifiers and dispersants. Solvents/auxiliaries suitablefor this purpose are essentially:

-   -   water, aromatic solvents (for example Solvesso® products,        xylene), paraffins (for example mineral oil fractions), alcohols        (for example methanol, butanol, pentanol, benzyl alcohol),        ketones (for example cyclohexanone, gamma-butyrolactone),        pyrrolidones (N-methylpyrrolidone, N-octylpyrrolidone), acetates        (glycol diacetate), glycols, fatty acid dimethylamides, fatty        acids and fatty acid esters. In principle, solvent mixtures may        also be used.    -   carriers such as ground natural minerals (for example kaolins,        clays, talc, chalk) and ground synthetic minerals (for example        highly disperse silica, silicates); emulsifiers such as        nonionogenic and anionic emulsifiers (for example        polyoxyethylene fatty alcohol ethers, alkylsulfonates and        arylsulfonates) and dispersants such as lignosulfite waste        liquors and methylcellulose.

Suitable surfactants used are alkali metal, alkaline earth metal andammonium salts of lignosulfonic acid, naphthalenesulfonic acid,phenolsulfonic acid, dibutylnaphthalenesulfonic acid,alkylarylsulfonates, alkyl sulfates, alkylsulfonates, fatty alcoholsulfates, fatty acids and sulfated fatty alcohol glycol ethers,furthermore condensates of sulfonated naphthalene and naphthalenederivatives with formaldehyde, condensates of naphthalene or ofnaphthalenesulfonic acid with phenol and formaldehyde, polyoxyethyleneoctylphenyl ether, ethoxylated isooctylphenol, octylphenol, nonylphenol,alkylphenyl polyglycol ethers, tributylphenyl polyglycol ether,tristearylphenyl polyglycol ether, alkylaryl polyether alcohols, alcoholand fatty alcohol ethylene oxide condensates, ethoxylated castor oil,polyoxyethylene alkyl ethers, ethoxylated polyoxypropylene, laurylalcohol polyglycol ether acetal, sorbitol esters, lignosulfite wasteliquors and methylcellulose.

Substances which are suitable for the preparation of directly sprayablesolutions, emulsions, pastes or oil dispersions are mineral oilfractions of medium to high boiling point, such as kerosene or dieseloil, furthermore coal tar oils and oils of vegetable or animal origin,aliphatic, cyclic and aromatic hydrocarbons, for example toluene,xylene, paraffin, tetrahydronaphthalene, alkylated naphthalenes or theirderivatives, methanol, ethanol, propanol, butanol, cyclohexanol,cyclohexanone, isophorone, highly polar solvents, for example dimethylsulfoxide, N-methylpyrrolidone and water.

Powders, materials for spreading and dustable products can be preparedby mixing or concomitantly grinding the active substances with a solidcarrier.

Granules, for example coated granules, impregnated granules andhomogeneous granules, can be prepared by binding the active compounds tosolid carriers. Examples of solid carriers are mineral earths such assilica gels, silicates, talc, kaolin, attaclay, limestone, lime, chalk,bole, loess, clay, dolomite, diatomaceous earth, calcium sulfate,magnesium sulfate, magnesium oxide, ground synthetic materials,fertilizers, such as, for example, ammonium sulfate, ammonium phosphate,ammonium nitrate, ureas, and products of vegetable origin, such ascereal meal, tree bark meal, wood meal and nutshell meal, cellulosepowders and other solid carriers.

In order to achieve good dispersion and adhesion of compositions withinthe present invention, it may be advantageous to formulate the wholebroth culture, supernatant and/or metabolite with components that aiddispersion and adhesion.

In general, the formulations comprise from 0.01 to 95% by weight,preferably from 0.1 to 90% by weight, of the components.

The chemical component (II) is employed in a purity of from 90% to 100%,preferably 95% to 100% (according to NMR spectrum).

The following are examples of formulations:

1. Products for Dilution with Water

A) Water-Soluble Concentrates (SL)

10 parts by weight of a composition according to the invention aredissolved in 90 parts by weight of water or in a water-soluble solvent.As an alternative, wetting agents or other auxiliaries are added.Dilution with water results in a formulation having a content of 10% byweight of components (I) and (II) is obtained.

B) Dispersible Concentrates (DC)

20 parts by weight of a composition according to the invention aredissolved in 70 parts by weight of cyclohexanone with addition of 10parts by weight of a dispersant, for example polyvinylpyrrolidone.Dilution with water gives a dispersion having a content of 0% by weightof components (I) and (II).

C) Emulsifiable Concentrates (EC)

15 parts by weight of a composition according to the invention aredissolved in 75 parts by weight of xylene with addition of calciumdodecylbenzenesulfonate and castor oil ethoxylate (in each case 5 partsby weight). Dilution with water gives an emulsion. The formulation has acontent of 15% by weight of components (I) and (II).

D) Emulsions (EW, EO)

25 parts by weight of a composition according to the invention aredissolved in 35 parts by weight of xylene with addition of calciumdodecylbenzenesulfonate and castor oil ethoxylate (in each case 5 partsby weight). This composition is introduced into 30 parts by weight ofwater by means of an emulsifying machine (Ultraturrax) and made into ahomogeneous emulsion. Dilution with water gives an emulsion. Theformulation has a content of 25% by weight of components (I) and (II).

E) Suspensions (SC, OD)

In an agitated ball mill, 20 parts by weight of a composition accordingto the invention are comminuted with addition of 10 parts by weight ofdispersants and wetting agents and 70 parts by weight of water or anorganic solvent to give a fine suspension. Dilution with water gives astable suspension having a content of 20% by weight of components (I)and (II).

F) Water-Dispersible Granules and Water-Soluble Granules (WG, SG)

50 parts by weight of a composition according to the invention areground finely with addition of 50 parts by weight of dispersants andwetting agents and prepared as water-dispersible or water-solublegranules by means of technical appliances (for example extrusion, spraytower, fluidized bed). Dilution with water gives a stable dispersion orsolution having a content of 50% by weight of components (I) and (II).

G) Water-Dispersible Powders and Water-Soluble Powders (WP, SP)

75 parts by weight of a composition according to the invention areground in a rotor-stator mill with addition of 25 parts by weight ofdispersants, wetting agents and silica gel. Dilution with water gives astable dispersion or solution having a content of 75% by weight ofcomponents (I) and (II).

2. Products to be Applied Undiluted

H) Dustable Powders (DP)

5 parts by weight of a composition according to the invention are groundfinely and mixed intimately with 95 parts by weight of finely dividedkaolin. This gives a dustable product having a content of 5% by weightof components (I) and (II).

J) Granules (GR, FG, GG, MG)

0.5 part by weight of a composition according to the invention is groundfinely and associated with 99.5 parts by weight of carriers. Currentmethods are extrusion, spray-drying or the fluidized bed. This givesgranules to be applied undiluted having a content of 0.5% of weight ofcomponents (I) and (II).

K) ULV Solutions (UL)

10 parts by weight of a composition according to the invention aredissolved in 90 parts by weight of an organic solvent, for examplexylene. This gives a product to be applied undiluted having a compoundcontent of 10% by weight of components (I) and (II).

Components (I) and (II) can be used as such, in the form of theirformulations or the use forms prepared therefrom, for example in theform of directly sprayable solutions, powders, suspensions ordispersions, emulsions, oil dispersions, pastes, dustable products,materials for spreading, or granules, by means of spraying, atomizing,dusting, spreading or pouring. The use forms depend entirely on theintended purposes; they are intended to ensure in each case the finestpossible distribution of components (I) and (II) according to theinvention.

Aqueous use forms can be prepared from emulsion concentrates, pastes orwettable powders (sprayable powders, oil dispersions) by adding water.To prepare emulsions, pastes or oil dispersions, the substances, as suchor dissolved in an oil or solvent, can be homogenized in water by meansof a wetting agent, tackifier, dispersant or emulsifier. However, it isalso possible to prepare concentrates composed of active substance,wetting agent, tackifier, dispersant or emulsifier and, if appropriate,solvent or oil, and such concentrates are suitable for dilution withwater.

The concentrations of the components in the ready-to-use preparationscan be varied within relatively wide ranges. In general, they are from0.0001 to 100%, preferably from 0.01 to 100%.

Components (I) and (II) may also be used successfully in theultra-low-volume process (ULV), it being possible to apply formulationscomprising over 95% by weight of active compound, or even to applycomponents (I) and (II) without additives.

Oils of various types, sun protectants, wetting agents or adjuvants maybe added to the component (I) or (II), even, if appropriate, not untilimmediately prior to use (tank mix). These agents are typically admixedwith component a) or b) according to the invention in a weight ratio offrom 1:100 to 100:1, preferably from 1:10 to 10:1.

In one embodiment, component (I) is applied together with a sunprotectant. Suitable sun protectants are, for example iron oxide ororganic UV photoprotective filters.

Organic UV photoprotective filters are understood as meaning organicsubstances which are able to absorb ultraviolet rays and give off theabsorbed energy again in the form of longer-wave radiation, e.g. heat.The term “Organic UV photoprotective filter” relates to one type or amixture of different types of said compounds. The organic substances maybe oil-soluble or water-soluble or they may be bound to a polymer. Thephotoprotective filters may be UV-A and/or UV-B filters, preferably UV-Bfilters.

UV-B filters which may be used are, for example, the followingsubstances:

-   -   3-benzylidenecamphor and derivatives thereof, e.g.        3-(4-methylbenzylidene)camphor;    -   4-aminobenzoic acid derivatives, preferably 2-ethylhexyl        4-(dimethylamino)benzoate, 2-octyl 4-(dimethylamino)benzoate and        amyl 4-(dimethylamino)benzoate;    -   esters of cinnamic acid, preferably 2-ethylhexyl        4-methoxycinnamate, propyl 4-methoxycinnamate, isoamyl        4-methoxycinnamate, isopentyl 4-methoxycinnamate, 2-ethylhexyl        2-cyano-3-phenylcinnamate (otocrylene);    -   esters of salicylic acid, preferably 2-ethylhexyl salicylate,        4-isopropylbenzyl salicylate, homomenthyl salicylate;    -   derivatives of benzophenone, preferably        2-hydroxy-4-methoxybenzophenone,        2-hydroxy-4-methoxy-4′-methylbenzophenone,        2,2′-dihydroxy-4-methoxybenzophenone;    -   esters of benzalmalonic acid, preferably 2-ethylhexyl        4-methoxybenzmalonate;    -   triazine derivatives, such as        2,4,6-trianilino(p-carbo-2′-ethyl-1′-hexyloxy)-1,3,5-triazine        (octyltriazone) and dioctylbutamidotriazone (Uvasorb® HEB).    -   Propane-1,3-diones, such as, for example,        1-(4-tert-butylphenyl)-3-(4′-methoxyphenyl)propane-1,3-dione.    -   2-phenylbenzimidazole-5-sulfonic acid and alkali metal, alkaline        earth metal, ammonium, alkylammonium, alkanolammonium and        glucammonium salts thereof;    -   sulfonic acid derivatives of benzophenones, preferably        2-hydroxy-4-methoxybenzophenone-5-sulfonic acid and its salts;    -   sulfonic acid derivatives of 3-benzylidenecamphor, such as, for        example, 4-(2-oxo-3-bornylidenemethyl)benzenesulfonic acid and        2-methyl-5-(2-oxo-3-bornylidene)sulfonic acid and salts thereof.

Preferred UV-B filters are derivatives of benzophenone.

Suitable UV-A filters are:

-   -   derivatives of benzoylmethane, for example        1-(4′-tert-butylphenyl)-3-(4′-methoxy-phenyl)propane-1,3-dione,        4-tert-butyl-4′-methoxydibenzoylmethane or        1-phenyl-3-(4′-isopropylphenyl)propane-1,3-dione;    -   Aminohydroxy-substituited derivatives of benzophenones, for        example N,N-diethylaminohydroxybenzoyl-n-hexylbenzoate.

Suitable adjuvants in this sense are in particular: organically modifiedpolysiloxanes, for example Break Thru S 240®; alcohol alkoxylates, forexample Atplus 245®, Atplus MBA 1303®, Plurafac LF 300® and Lutensol ON30®; EO/PO block polymers, for example Pluronic RPE 2035® and GenapolB®; alcohol ethoxylates, for example Lutensol XP 80®; and sodiumdioctylsulfosuccinate, for example Leophen RA®.

For seed treatment purposes, respective formulations can in certaincases be diluted 2-10 fold leading to concentrations in the ready to usepreparations of 0.01 to 60% by weight active compound by weight,preferably 0.1 to 40% by weight.

Conventional seed treatment formulations include for example flowableconcentrates FS, solutions LS, powders for dry treatment DS, waterdispersible powders for slurry treatment WS, water-soluble powders SSand emulsion ES and EC and gel formulation GF. These formulations can beapplied to the seed diluted or undiluted. Application to the seeds iscarried out before sowing, either directly on the seeds.

In one embodiment a FS formulation is used for seed treatment.Typcially, a FS formulation may comprise 1-800 g/l of active ingredient,1-200 g/l surfactant, 0 to 200 g/l anti-freezing agent, 0 to 400 g/l ofbinder, 0 to 200 g/l of a pigment and up to 1 liter of a solvent,preferably water.

In accordance with one variant of the present invention, a furthersubject of the invention is a method of treating soil by theapplication, in particular into the seed drill: either of a granularformulation containing the plant health composition of the invention—incombination or as a composition/formulation, or of a mixture of twogranular formulations, each containing one of the two activeingredients, with optionally one or more solid or liquid, agriculturallyacceptable carriers and/or optionally with one or more agriculturallyacceptable surfactants. This method is advantageously employed inseed-beds of cereal, maize, cotton and sunflower. The rates for eachactive ingredient may be in the range of 10 to 1000 g/ha, as for example50 to 500 g/ha or 50 to 200 g/ha.

The seed treatment application is carried out by spraying or dusting theseeds before sowing of the plants and before emergence of the plants.

In the treatment of seeds the corresponding formulations are applied bytreating the seeds with an effective amount of component (I) andoptionally at least one component (II). Herein, the application rates ofthe composition of the invention are generally from 0.1 g to 10 kg per100 kg of seed, preferably from 1 g to 5 kg per 100 kg of seed, inparticular from 1 g to 2.5 kg per 100 kg of seed. For specific cropssuch as lettuce the rate can be higher. In seed treatment applicationsin which component (I) is derived from a whole broth of the Bacillussubtilis strain or its mutants the number of colony forming units (CFU)applied is important and is generally from 1×10⁸ through 1×10¹² peracre, preferably from 1×10⁹ through 1×10¹³ per acre or, in particularfrom 1×10¹⁰ through 1×10¹² per acre.

The following examples shall illustrate the invention without limitingit. All plants in the experiments listed below, could grow underessentially non-existent pathogen pressure.

EXAMPLE 1 Tomato

Bacillus subtilis QST 713 was used to treat tomato seeds via aquasi-soil drench in the greenhouse. Specifically, tomato seeds wereplanted in steam-sterilized trays containing sterile media and grown inthe greenhouse using standard techniques. Serenade® ASO, which is aliquid formulation of Bacillus subtilis QST 713 containing 1×10⁹ CFU(Colony Forming Units)/g was applied to media at rates of 4 oz, 8 oz,and 16 oz per acre once, at the time of planting. The Serenade® ASOproduct was applied as a spray application and not as a true drench, asthe spray application did not provide enough water to cause germination.Other seeds not treated with the Serenade® ASO product were used asnegative controls.

At the time of transplanting to the field, plants grown in media treatedwith Serenade® ASO showed a higher vigor than those in the untreatedcontrol group based on grower observations of plant height, size of rootmass as well as color and spindliness of leaves. In addition, a higherpercentage of plants treated with the Serenade® ASO product were able tobe used in the field than those in the untreated control group (table1a).

TABLE 1a Plants that were able to Treatment be used in the field (%)Control 80.3 Serenade ® ASO 4 oz 83.0 Serenade ® ASO 8 oz 88.3Serenade ® ASO 16 oz 87.7

As can be seen in table 1a, Serenade ASO has a positive effect on planthealth by increasing the plant's vigor resulting in an increased numberof plants that could be used for planting in the field.

Useable transplants treated with Serenade ASO and controls weresubsequently planted in the field and grown under the same standardconditions (with all receiving the same watering, pesticideapplications, and the like) until harvest. Due to the pesticideapplication, the plants could grow under essentially non-existentpathogen pressure. At harvest, plants treated with Serenade ASO at thetime of planting in the greenhouse yielded more total weight of tomatoesand more marketable tomatoes than the untreated control (table 1b).

TABLE 1b Yield (Total weight of Marketable tomatoes in 12 plots,tomatoes Treatment each containing 2 plants) (%) Control 359 46Serenade ® ASO 4 oz/ac 366 78 Serenade ® ASO 8 oz/ac 397 71 Serenade ®ASO 16 oz/ac 368 77

As can be seen in table 1 b, Serenade® ASO has also a positive effect onplant health by increasing the plant's yield (total weight of tomatoes).In addition, the treatment with Serenade® ASO results in increased vigorof the plants and cosequently in more marketable tomatoes compared tothe untreated control plants.

EXAMPLE 2 Pepper

Bacillus subtilis QST 713 was used to treat pepper seeds via aquasi-soil drench in the greenhouse. Specifically, pepper seeds wereplanted in steam-sterilized trays containing sterile media and grown inthe greenhouse using standard techniques. Serenade® ASO, which is aliquid formulation of Bacillus subtilis QST 713 containing 1×109 CFU/g,was applied to media at rates of 4 oz, 8 oz, and 16 oz per acre once, atthe time of planting. The Serenade® ASO product was applied as a sprayapplication and not as a true drench, as the spray application did notprovide enough water to cause germination. Other seeds not treated withthe Serenade® ASO product were used as negative controls.

At the time of transplanting to the field, plants grown in media treatedwith Serenade® ASO showed a higher vigor than those in the untreatedcontrol group based on grower observations of plant height, size of rootmass as well as color and spindliness of leaves. In addition, a higherpercentage of plants treated with the Serenade® ASO product were able tobe used in the field compared to those in the untreated control group(table 2).

TABLE 2 Vigor (0 = no vigor; 10 = Plants that were able to Treatmentoptimal vigor) be used in the field (%) Control 3.7 95.3 Serenade ® ASO4 oz/ac 4.0 96.0 Serenade ® ASO 8 oz/ac 5.3 97.0 Serenade ® ASO 16 oz/ac6.7 97.0

As can be seen in table 2, Serenade® ASO has a positive effect on planthealth by increasing the plant's vigor. In addition, the treatment withSerenade® ASO results in more plants that were able to be used in thefield compared to the untreated control plants which in turn will resultin an increased overall yield.

EXAMPLE 3 Broccoli

Bacillus subtilis QST 713 was used to treat broccoli seeds via aquasi-soil drench in the greenhouse. Specifically, broccoli seeds wereplanted in steam-sterilized trays containing sterile vermiculite andgrown in the greenhouse using standard techniques. Serenade® ASO, whichis a liquid formulation of Bacillus subtilis QST 713 containing 1×10⁹CFU/g, was applied to media at rates of 4 oz, 8 oz, and 16 oz per acreonce, at the time of planting. The Serenade® ASO product was applied asa spray application and not as a true drench, as the spray applicationdid not provide enough water to cause germination. Other seeds nottreated with the Serenade® ASO product were used as negative controls.

At the time of transplanting to the field, plants grown in media treatedwith Serenade® ASO showed a higher vigor than those in the untreatedcontrol group based on grower observations of plant height, size of rootmass, as well as color and spindliness of leaves. In addition, a higherpercentage of plants treated with the Serenade® ASO product were able tobe used in the field compared to those in the untreated control group(table 3).

As can be seen in table 3, Serenade® ASO has a positive effect on planthealth by increasing the plant's vigor. In addition, the treatment withSerenade® ASO results in more plants that were able to be used in thefield compared to the untreated control plants which in turn will resultin an increased overall yield.

TABLE 3 Vigor (0 = no vigor; 10 = Plants that were able to Treatmentoptimal vigor) be used in the field (%) Control 4.7 91.7 Serenade ® ASO4 oz/ac 6.0 92.0 Serenade ® ASO 8 oz/ac 7.3 93.0 Serenade ® ASO 16 oz/ac5.3 93.0

EXAMPLE 4 Wheat

Wheat seed was treated with Bacillus subtilis QST 713 by applying to theseeds a slurry of the Serenade® ASO product at a rate of 4 oz, 8 oz, 12oz or 16 oz per 100 lb seed. The slurry was prepared by mixing Serenade®ASO with water. Seeds remained in the slurry for various periods oftime, ranging from overnight to two weeks. Fields were seeded at a rateof 80-100 lb per acre. Seeds were applied to fields in which diseasepressure was essentially non-existent. Consequently, growers wouldtypically not engage in seed treatment for disease control.

TABLE 4 Yield (bushels/ Treatment acre) Control 49.9 Difenoconazole +mefenoxam 65.7 Serenade ® ASO 4 oz + difenoconazole + mefenoxam 79.5Serenade ® ASO 4 oz/100 lb seed 100.4 Serenade ® ASO 8 oz/100 lb seed90.8 Serenade ® ASO 12 oz/100 lb seed 49.3 Serenade ® ASO 16 oz/100 lbseed 34.6

As can be seen in table 4, Serenade® ASO has a very positive effect onplant health by increasing the yield when applied below 10 oz/100 lbseed. When applied at higher amounts with this particular formulation,the yield may remain unaffected or may even decline. However, it isunknown whether this decline is due to a formulation inert in thisparticular formulation or due to the rate of active ingredient. One ofordinary skill in the art would be able to determine the optimal rate ofapplication of component (I) with routine experimentation.

EXAMPLE 5 Wheat

The Serenade® ASO product, which contains 1×109 CFU/g Bacillus subtilisQST 713 was applied in furrow at the time of wheat seed planting alongwith the following starter fertilizer: 10-34-0 (10% nitrogen, 34%phosphate and 0% potassium) and/or Power Up (6% nitrogen, 18% phosphateand 6% potassium) at the rates/per acre shown below. Disease pressurewas essentially non-existent, such that disease rates were not reportedfor this trial. This was a situation in which a grower would nottypically apply the Serenade® ASO product, as the cost would not bejustified from a disease control perspective.

TABLE 5 Yield (bushels/ Treatment acre) Control 21.7 3 gallons per acre(gpa) of 10-34-0 25.9 2 gpa of 10-34-0 + 1 gpa Power Up 28.3 2 gpa of10-34-0 + 1 gpa Power Up + 31.0 Serenade ® ASO 8 oz/ac 2 gpa of10-34-0 + 1 gpa Power Up + 30.1 Serenade ® ASO 16 oz/ac

As can be seen in table 5, the application of Serenade® ASO with thefertilizers resulted in an increased yield. In addition, in the case ofin-furrow application of Serenade® ASO, wheat heads displayed advancedmaturity compared to the untreated control plants in which only 75% ofthe wheat heads had formed at the same time point.

EXAMPLE 6 Lettuce

The active compounds were used applying commercially availableformulations and diluted according to the concentrations/dose rates asstated in table 6. Commercially available lettuce seedlings(“Eichblatt”) were used for the described greenhouse trial. 4replications (pots with 1 plant each) were used per treatment. Plantswere grown in commercially available substrate (Floradur A) at approx.20° C. in the greenhouse. Drench applications using a volume of 25 ml ofproduct solution or water (Control) were made on 16 consecutive days. Onthe last day, fresh weight was determined using all plant parts aboveground.

TABLE 6 Yield Fresh Treatment weight (g) Control 51.7 Serenade ® MAX 312ppm 59.5 Serenade ® MAX 625 ppm 62.4 Serenade ® MAX 1250 ppm 74.1Serenade ® MAX 2500 ppm 68.9

As can be seen from table 6, Serenade® MAX strongly increases the freshweight of lettuce plants which is an essential parameter for vegetables.

EXAMPLE 7 Soybeans

Soybeans were planted in December 2008 at the BASF experimental stationin Campinas, San Antonio de Posse, Sao Paulo, Brazil. The varietyEmprapa 48 was planted at a seeding rate of 300,000 plants per ha. Rowspacing was 45 cm. Two trials were setup as a randomized bloc designwith 6 replications. Plot size was 20 m².

Bacillus subtilis QST 713 was applied by foliar application to thevegetative parts of the soybean plants at developmental stage 23/29(BBCH) followed by the foliar application of either Bacillus subtilisQST 713 alone or in tank mix with pyraclostrobin (applied as COMET® atthe beginning of flowering at the developmental stages 60/63 (BBCH). Theactive ingredients were applied using the commercial formulationsSerenade® (10%, WP with 5×109 cfu/g) and Comet® (250 g/L, EC). Theformulations were used in the dose rates given in table 7. Total sprayvolume for foliar applications was 150 l/ha. Serenade® was applied with3 kg product per ha and Comet® applied with a product rate of 0.4 l/ha.At maturity the crop was harvested and grain yield was measured in t/ha.Green leaf area was assessed 31 days after the last treatment (table 7)by estimating the green leaf area in 10 randomly chosen plants per plot.

The efficacy (E) was calculated as % increase of green leaf area in thetreatments compared to the untreated control according to the followingformula:

E=a/b−1·100

-   -   E efficacy    -   a corresponds to the green leaf area (%) of the treated plants        and    -   b corresponds to the green leaf area (%) of the untreated        (control) plants

An efficacy (E) of 0 means the green leaf area of the treated plantscorresponds to that of the untreated control plants; an efficacy of 100means the treated plants showed an increase in the green leaf area of100%.

TABLE 7 Grain GLA GLAE Yield Treatment PR FC FT AT (%) (%) (t/ha)Untreated 7.5 1.97 Serenade ® 3.0 kg/ha 10% WP 23/29 12.5 66.6 2.30 3.0kg/ha 10% WP 60/63 Serenade ® 3.0 kg/ha 10% WP 23/29 20.0 166.6 3.08 3.0kg/ha 10% WP 60/63 Pyra- 0.3 l/ha 250 g/l EC 60/63 clostrobin PR =Product rate; FC = Formulation concentration; FT = Formulation type; AT= Application time (BBCH); GLA = Green Leaf Area; Green Leaf Area(Efficacy)

As can be seen in table 7, Serenade® clearly increases the green leafarea duration (maintanance of green leaves) and the grain yield insoybeans compared to the untreated control. In addition, the resultsshown in table 7 demonstrate that the efficacy of the combination ofSerenade® and pyraclostrobin is even higher than for Serenade alone. Anincrease of the green leaf area is a visible sign of the enhanced plantvigor. Based on a prolonged maintanance of green leaves which in turnresults in a prolonged photosynethic activity of the leaves as well asan overall srengthening of the plant, the plant is able to produce ahigher yield.

EXAMPLE 8 Soybeans

Soybeans were planted in 2009 at 9 locations across the soybean growingarea of Midwest of the USA (IN, IL, IA, MO, NE, and SD). Planting datesranched from May 7th at York, Neb., to June 22nd at Clarence, Mo.Bacillus subtilis QST 713 was applied to the vegetative parts of thesoybean plants at developmental stage 23/29 (BBCH). The B. subtilis QST713 was applied using the commercial formulation Serenade Max® (14.3%,WP with 7.3×109 cfu/g). Serenade Max® was used in the dose rates givenin Table 8. Total spray volume for foliar application ranched from 140to 200 l/ha. Serenade Max® was applied at 3 kg product per ha. Atmaturity the crop was harvested and grain yield was measured as t/ha(table 8). Green leaf area was assessed 36 to 66 days after the lasttreatment on seven of the trial locations (table 8) by estimating thegreen leaf area in 10 randomly chosen plants per plot. The efficacy wascalculated as indicated above.

TABLE 8 Grain GLA GLAE Yield Treatment PR FC FT AT (%) (%) (t/ha)Untreated 64.36 34.87 Serenade ® 3.0 kg/ha 14.3% WP 23/29 67.04 4.236.42 MAX PR = Product rate; FC = Formulation concentration; FT =Formulation type; AT = Application time (BBCH); GLA = Green Leaf Area;Green Leaf Area (Efficacy)

As can be seen in table 8, Serenade Max® increases the green leaf areaduration and therefore improve photosynthetic activity of soybeans. Inaddition, Serenade Max® strongly increases the grain yield; in this caseby 1.55 t/ha in soybeans compared to the untreated control by improvingthe vigor of the soybean plants.

EXAMPLE 9 Winter Wheat

Winter wheat was grown in the 2008/2009 growing season at 4 locationsacross Germany (Thuringia, Baden-Wuerttemberg, andRhineland-Palatinate). Seeding of the crop ranged from September 21st toOctober 26th. The trials were setup in a randomized bloc design with 6replications. Bacillus subtilis QST 713 was applied to the winter wheatplants at beginning of shooting (growth stage 31/32, BBCH). Thefungicide spray sequence consisted of an application of epoxiconazole atbeginning of shooting followed by an application of epoxiconazole incombination with pyraclostrobin at flag leaf stage (growth stage 37/39).The B. subtilis QST 713 was applied using the commercial formulationSerenade Max® (14.3%, WP with 7.3×109 cfu/g). Epoxiconazole was appliedalone at beginning of shooting as the commercially available formulationOpus® (125 g/l, SC). The combination of epoxiconazole and pyraclostrobinwas applied as a ready to use developmental Opera® formulation (SE)containing 62.5 g/l epoxiconazole and 85 g/l pyraclostrobin. Productrates are given in table 9. Total spray volume for the foliarapplications were 300 l/ha. At maturity the crop was harvested and grainyield was measured as t/ha (table 9).

TABLE 9 Observed Grain yield Yield increase Treatment PR FC FT AT (t/ha)(t/ha) Untreated 6.73 Epoxiconazole 0.8 l/ha 125 g/l SC 31/32 7.78 1.05Epoxiconazole + 2.0 l/ha 147.5 SE 37/39 Pyraclostrobin Serenade 3.0kg/ha 14.3% WP 31/32 6.87 0.14 Max ® Serenade 3.0 kg/ha 14.3% WP 31/327.93 1.20 Max ® Epoxiconazole 0.8 l/ha 125 g/l SC 31/32 Epoxiconazole +2.0 l/ha 147.5 SE 37/39 Pyraclostrobin PR = Product rate; FC =Formulation concentration; FT = Formulation type; AT = Application time(BBCH)

As can be seen in table 9, the Serenade Max® treatment increased theyield of the wheat crop by 140 kg/ha. The joint application with anapplication sequence of Serenade Max® together with the fungicide sprayof epoxiconazole and epoxiconazole plus pyraclostrobin was even superiorcompared to the application of the fungicides (epoxiconazole pluspyraclostrobin) alone (1.2 vs. 1.05 t/ha). These findings demonstratethe improvement of the vigor of the wheat plants by Bacillus subtilisQST 713 and the superior effect of the combination of Bacillus subtilisQST 713 with fungicides compared to the effect of the solo applicationof Bacillus subtilis QST 713 or an application of a compositioncomprising as active ingredients only fungicides.

EXAMPLE 10 Maize (Corn)

Maize was planted in 2009 at Carlyle, Ill., USA. The variety Burrus616XLR was planted at a conventional seeding rate and a row spacing of76 cm. The trial was setup as a randomized bloc design with 6replications. Plot size was 18 m². Pyraclostrobin was applied atdevelopmental stage 34/37 (BBCH). Bacillus subtilis QST 713 (SerenadeMax®) was applied to the maize plants at developmental stage 34/37(BBCH) followed by a second application at developmental stage 55/57(BBCH). Bacillus subtilis QST 713 (Serenade Max®) in combination withpyraclostrobin were applied as a tank mix at the developmental stage34/37 (BBCH). The active ingredients were applied using commercialformulations Serenade Max® (14.3%, WP with 7.3×109 cfu/g) and Headline®(250 g/L, EC). The formulations were used in the dose rates given intable 10. Total spray volume for foliar applications was 200 l/ha.Serenade Max® was applied with 2.1 kg product per ha and Headline® witha product rate of 0.44 l/ha. At maturity the crop was harvested andgrain yield was measured as t/ha (table 10).

The expected yield increase by the combination of the active compoundswas estimated using Colby's formula (Colby, S. R., Calculatingsynergistic and antagonistic responses of herbicide combinations, Weeds,15, pp. 20-22, 1967) and compared with the observed yield increase.

E=x+y−x·y/100   Colby's formula:

-   -   E expected efficacy, expressed as the numerical difference of        the yield in t/ha to the untreated control, when using the        mixture of the active compounds A and B at the concentrations a        and b    -   x efficacy, expressed as the numerical difference of the yield        in t/ha to the untreated control, when using the active        ingredient A at the concentration a    -   y efficacy, expressed as the numerical difference of the yield        in t/ha to the untreated control, when using the active        ingredient B at the concentration b

TABLE 10 Observed Expected Grain yield yield Yield increase increaseTreatment PR FC FT AT (t/ha) (t/ha) (t/ha) Untreated 8.27 Pyraclostrobin0.44 l/ha 250 g/l EC 34/37 8.32 0.05 Serenade Max ® 3.0 kg/ha 14.3% WP34/37 8.42 0.15 3.0 kg/ha 55/57 Serenade Max ® 3.0 kg/ha 14.3% WP 34/378.66 0.39 0.19 Pyraclostrobin 0.3 l/ha 250 g/l EC PR = Product rate; FC= Formulation concentration; FT = Formulation type; AT = Applicationtime (BBCH)

Application of Serenade Max® alone and the combination of Serenade Max®and pyraclostrobin results in a clear yield increase. Compared to theyield increase of the solo application of Serenade Max® orpyraclostrobin solo, the yield increase when the combination of SerenadeMax® and pyraclostrobin were applied togehter is even higher than couldhave been expected according to Colby's formula. This yield increase,which is about double as high as expected, clearly demonstrates thesynergistic effect of the compositions according to the invention on theplant's vigor and the plant's yield.

EXAMPLE 11 Winter Wheat

Winter wheat was grown in the 2008/2009 growing season at Cagnicourt inFrance. The variety Premio was sown November 1st at a seeding rate of125 kg/ha. The trial was setup in a randomized bloc design with 6replications and a plot size of 22.5 m² . Bacillus subtilis QST 713 wasapplied to the winter wheat plants at beginning of shooting (growthstage 31/32, BBCH). The fungicide spray sequence consisted of anapplication of epoxiconazole at beginning of shooting followed by anapplication of epoxiconazole in combination with pyraclostrobin at flagleaf stage (growth stage 37/39). The B. subtilis QST 713 was appliedusing the commercial formulation Serenade Max® (14.3%, WP with 7.3×109cfu/g). Epoxiconazole was applied alone at beginning of shooting as thecommercially available formulation Opus® (125 g/l, SC). The combinationof epoxiconazole and pyraclostrobin was applied as a ready to usedevelopmental Opera formulation (SE) containing 62.5 g/l epoxiconazoleand 85 g/l pyraclostrobin. Product rates are given in table 11. Totalspray volume for the foliar applications were 300 l/ha. At end ofheading and beginning of flowering, respectively, the number ears per m²was counted (table 11). The expected increase in number of ears per m²by the combination of the active compounds was estimated using Colby'sformula (Colby, S. R., Calculating synergistic and antagonisticresponses of herbicide combinations, Weeds, 15, pp. 20-22, 1967) andcompared with the observed increase as described above.

TABLE 11 No. of Observed Expected Treatment PR FC FT AT Ears/m² increaseincrease Untreated 375 Epoxiconazole 0.8 l/ha 125 g/l SC 31/32 394 19Epoxiconazole + 2.0 l/ha 147.5 SE 37/39 Pyraclostrobin Serenade Max ®3.0 kg/ha 14.3% WP 31/32 385 10 Serenade Max ® 3.0 kg/ha 14.3% WP 31/32Epoxiconazole 0.8 l/ha 125 g/l SC 31/32 406 31 27 Epoxiconazole + 2.0l/ha 147.5 SE 37/39 Pyraclostrobin PR = Product rate; FC = Formulationconcentration; FT = Formulation type; AT = Application time (BBCH)

In this example, again, Serenade Max® improved the health of the wheatplants leading to increased number of ears per m². An increased numberof ears per m² was also observed from the fungicide spray sequence. Theincrease observed from the combined application of Serenade Max® and thefungicides was higher than expected according to Colby's formula, as isshown in table 11. This result clearly illustrates the synergisticeffect of the combination of Bacillus subtilis QST 713 with azoles andstrobilurins on the vigor and yield of the wheat plants when appliedtogether in a tank mix or a spray sequence.

EXAMPLE 12 Peas

The active compounds were used applying commercially availableformulations and diluted according to the concentrations/dose rates asstated in the respective data tables.

Seed treatment was done to seeds of peas using the Hege Seed Treatmentequipment in a sequential approach. Pyraclostrobin was applied with avolume of 850 ml slurry per 100 kg seeds. Afterwards the stated amountof Serenade® MAX was dissolved in a total volume of 8.3 L water (amountfor 100 kg seeds) and applied sequentially in ten steps with drying ofthe seeds inbetween. The compounds were used as commercial finishedformulations and diluted with water to the stated concentration of theactive compound.

Seeds of peas were sown in soil (10 seeds/pot, 10replications/treatment) and incubated in the greenhouse at 20° C. for 12days. Plants were harvested and pooled per treatment and the plant freshweight was determined.

The expected plant fresh weights of active compound mixtures weredetermined using Colby's formula as defined above [R. S. Colby,“Calculating synergistic and antagonistic responses of herbicidecombinations”, Weeds 15, 20-22 (1967)] and compared with the observedplant fresh weights.

As can be seen in table 12, the measured fresh weight when Serenade Max®is applied strongly increases compared to the untreated control plants.In case a mixture according to the present invention is applied such asthe combination of Serenade Max® and pyraclostrobin, the fresh weight asan indicator for the plant's vigor and yield is increased evensynergistically.

TABLE 12 Dose rate Fresh Calculated (g/100 weight efficacy accordingTreatment kg seed) (g) to Colby (%) Untreated 55 Pyraclostrobin 5 47(200 g/L, FS) Serenade Max ® 173 64 (14.3% B. subtilis, WP)Pyraclostrobin + 5 92 81 Serenade Max ® 173

1. A method for increasing the vigor and/or crop yield of agriculturalplants under essentially non-existent pathogen pressure, wherein theplants, the plant propagules, the seed of the plants and/or the locuswere the plants are growing or are intended to grow are treated with aneffective amount of a composition comprising the Bacillus subtilisstrain with NRRL Accession No. B-21661 or a cell-free, extract thereof,and/or a mutant of this strain or extract having all the identifyingcharacteristics of the respective strain or extract.
 2. The method asclaimed in claim 1, wherein a commercially available formulation of theBacillus subtilis strain is used.
 3. The method as claimed in claim 1,further comprising treating the plants, the plant propagules, the seedof the plants and/or the locus where the plants are growing or areintended to grow with an effective amount of at least one chemicalcompound selected from the active compound groups A) to J): A)strobilurins selected from the group consisting of azoxystrobin,dimoxy-strobin, enestroburin, fluoxastrobin kresoxim-methyl,metomi-nostrobin, orysastrobin, picoxystrobin, pyraclostrobin,pyribencarb, trifloxystrobin,2-(2-(6-(3-chloro-2-methyl-phenoxy)-5-fluoro-pyrimidin-4-yloxy)-phenyl)-2-methoxyimino-N-methyl-acetamide,3-methoxy-2-(2-(N-(4-methoxy-phenyl)-cyclopropane-carboximidoyl-sulfanylmethyl)-phenyl)-acrylicacid methyl ester, methyl(2-chloro-5-[1-(3-methylbenzyloxvimino)ethyl]benzyl)carbamate and2-(2-(3-(2,6-dichlorophenyl)-1-methyl-allylideneaminooxymethyl)-phenyl)-2-methoxylmino-N-methyl-acetamide;B) carboxamides selected from the group consisting of carboxanilides:benalaxyl, benalaxyl-M, benodanil, bixafen, boscalid, carboxin,fenfuram, fenhexamid, flutolanil, furametpyr, isopyrazam, isotianil,kiralaxyl, mepronil, metalaxyl, metalaxyl-M (mefenoxam), ofurace,oxadixyl, oxycarboxin, penthiopyrad, sedaxane, tecloftalam,thifluzamide, 2-amino-4-methyl-thiazole-5-carboxanilide,2-chloro-N-(1,3-trimethyl-indan-4-yl)-nicotinamide,N-(3′,4′,5′-trifluorobiphenyl-2-yl)-3-diftuoromethyl-1-methyl-1H-pyrazole-4-carboxamide,(4′-trifluoromethylthiobiphenyl-2-yl)-3-difluorometh-1-methyl-1H-pyrazole-4-carboxamide,N-(2-(1,3-dimethyl-butyl)-phenyl)-1,3-dimethyl-5-fluoro-1H-pyrazole-4-carboxamideandN-(2-(1,3,3-trimethyl-butyl)-phenyl)-1,3-dimethyl-5-fluoro-1H-pyrazole-4-carboxamide;carboxylic morpholides: dimethomorph, flumorph, pyrimorph; benzoic acidamides: flumetover, fluopicolide, fluopyram, zoxamide,N-(3-Ethyl-3,5,5-trimethyl-cyclohexyl)-3-formylamino-2-hydroxy-benzamide;other carboxamides: carpropamid, dicyriomet, mandiproamid,oxytetracyclin, silthiofarm andN-(6-methoxy-pyridin-3-yl)cyclopropanecarboxylic acid amide; C) azolesselected from the group consisting of triazoles: azaconazole,bitertanol, bromuconazole, cyproconazole, difenoconazole, diniconazole,diniconazole-M, epoxiconazole, fen-buconazole, fluquinconazole,flusilazole, flutriafol, hexaconazole, imibenconazoie, ipconazole,metconazole, myclobutanil, oxpoconazole, paclobutrazole, penconazole,propiconazole, prothioconazole, simeconazole, tebuconazole,tetraconazole, triadimefon, triadimenol, triticonazole, uniconazole,1-(4-chloro-phenyl)-2-([1,2,4]triazol-1-yl)-cycloheptanol; imidazoles:cyazofamid, imazalil, pefurazoate, prochloraz, triflumizol;benzimidazoles: benomyl, carbendazim, fuberidazole, thiabendazole;others: ethaboxam, etridiazole, hymexazole and2-(4-chloro-phenyl)-N-[4-(3,4-dimethoxy-phenyl)-isoxazol-5-yl]-2-prop-2-ynyloxy-acetamide;D) heterocyclic compounds selected from the group consisting ofpyridines: fluazinam, pyrifenox,3-[5-(4-chloro-phenyl)-2,3-dimethyl-3-[5-(4-methyl-phenyl)-2,3-dimethyl-isoxazolidin-3-yl]]pyridine,2,3,5,6-tetra-chloro-4-methanesulfonyl-pyridine,3,4,5-trichloropyridine-2,6-di-carbonitrile,N-(1-(5-bromo-3-chloro-pyridin-2-yl)-ethyl)-2,4-dichloronicotinamide,N-[(5-bromo-3-chloro-pyridin-2-yl)-methyl]-2,4-dichloro-nicotinamide;pyrimidines: bupirimate, cyprodinil, diflumetorim, fenarimol, ferimzone,mepanipyrim, nitrapyrin, nuarimol, pyrimethanil; piperazines: triforine;pyrroles: fludioxonil; morpholines: aldimorph, dodemorph,dodemorph-acetate, fenpropimorph, tridemorph; piperidines: fenpropidin;dicarboximides: fluoroimid, iprodione, procymidone, vinclozolin;non-aromatic 5-membered heterocycles: famoxadone, fenamidone,octhilinone, probenazole,5-amino-2-isopropyl-3-oxo-4-ortho-tolyl-2,3-dihydro-pyrazole-1-carbothioicacid S-allyl ester; others: acibenzolar-S-methyl, amisulbrom, anilazin,blasticidin-S, captafol, captan, chinomethionat, dazomet, debacarb,diclomezine, difenzoquat, difenzoquat-methylsulfate, fenoxanil, Folpet,oxolinic acid, piperalin, proquinazid, pyroquilon, quinoxyfen,triazoxide, tricyclazole, 2butoxy-6-iodo-3-propylchromen-4-one,5-chloro-1-(4,6-dimethoxy-pyrimidin-2-yl)-2-methyl-1H-benzoimidazole,5-chloro-7-(4-methylpiperidin-1-yl)-6-(2,4,6-trifluorophenyl)-[1,2,4]triazolo-[1,5-a]pyrimidineand 5-ethyl-6-octyl-[1,2,4]triazolo[1,5-a]pyrimidine-7-ylamine; E)carbamates selected from the group consisting of thio- anddithiocarbamates: ferbam, mancozeb, maneb, metam, methasulphocarb,metiram, propineb, thiram, zineb, ziram; carbamates: benthiavalicarb,diethofencarb, iprovalicarb, propamocarb, propamocarb hydrochlorid,valiphenal and N-(1-(1-(4-cyano-phenyl)ethanesulfonyl)-but-2-yl)carbamicacid-(4-fluorophenyl)ester; F) other active, substances selected fromthe group consisting of guanidines: guanidine, dodine, dodine free base,guazatine, guazatine-acetate, iminoctadine, iminoctadine-triacetate,iminoctadine-tris(albesilate); antibiotics: kasuamycin, kasugamycinhydrochloride-hydrate, streptomycin, polyoxine, validamvcin A,streptomycin; nitrophenyl derivates: binapacryl, dinobuton, dinocap,nitrthal-isopropyl, tecnazen; organometal compounds: fentin salts, suchas fentin-acetate, fentin chloride or fentin hydroxide;sulfur-containing heterocyclyl compounds: dithianon, isoprothiolane;organophosphorus compounds: edifenphos, fosetyl, fosetyl-aluminum,iprobenfos, phosphorous acid and its salts, pyrazophos,tolclofos-methyl; organochlorine compounds: chlorothalonil,dichlofluanid, dichlorophen, flusulfamide, hexachlorobenzene, pencycuronpentachlor-phenole and its salts, phthalide, quintozene,thiophanate-methyl, tolylfluanid,N-(4-chloro-2-nitro-phenyl)-N-ethyl-4-methyl-benzenesulfonamide;inorganic active substances: Bordeaux mixture, copper acetate, copperhydroxide, copper oxychloride, basic copper sulfate, sulfur; others:biphenyl, bronopol, cyflufenamid, cymoxanil, diphenylamin, metrafenone,mildiomycin, oxin-copper, prohexadione-calcium, spiroxamine,tolylfluanid,N-(cyclopropylmethoxyimino-(6-difluoro-methoxy-2,3-difluoro-phenyl)-methyl)-2-phenylacetamide,N′-(4-chloro-3-trifluoromethyl-phenoxy)-2,5-dimethyl-phenyl)-N-ethyl-N-methylformamidine,N′-(4-(4-fluoro-3-trifluoromethyl-phenoxy)-2,5-dimethyl-phenyl)-N-ethyl-N-methylformamidine,N′-(2-methyl-5-trifluoromethyl-4-(3-trimethylsilanyl-propoxy)-phenyl)-N-ethyl-N-methylformamidine,N′-(5-difluoromethyl-2-methyl-4-(3-trimethysilanyl-propoxy)-phenyl)-N-ethyl-N-methylformamidine,2-{1-[2-(5-methyl-3-trifluoromethyl-pyrazole-1-yl)acetyl]-piperidin-4-yl}-thiazole-4-carboxylicacid methyl-(1,2,3,4-tetrahydro-naphthalen-1-yl)-amide,2-{1-[2-(5-methyl-3-trifluoromethyl-pyrazole-1-yl)acetyl]-piperidin-4-yl}-thiazole-4-carboxylicacid methyl-(R)-1,2,3,4-tetrahydro-naphthalen-1-yl-amide, acetic acid6-tert-butyl-8-fluoro-2,3-dimethyl-quinolin-4-yl ester andmethoxy-acetic acid 6-tert-butyl-8-fluoro-2,3-dimethyl-quinolin-4-ylester, fentin acetate, fentin chloride, fentin hydroxide; G) plantgrowth regulators (PGRs) selected from the group consisting of abscisicacid, amidochlor, ancymidol, 6-benzylaminopurine, brassinolide,butralin, chlormequat (chlormequat chloride), choline chloride,cyclanilide, daminozide, dikegulac, dimethipin, 2,6-dimethylpuridine,ethephon, flumetralin, flurprimidol, fluthiacet, forchlorfenuron,gibberellic acid, inabenfide, indole-3-acetic acid, maleic hydrazide,mefluidide, mepiquat (mepiquat chloride), naphthaleneacetic acid,N-6-benzyladenine, paclobutrazol, prohexadione (prohexadione-calcium),prohydrojasmon, thidiazuron, triapenthenol, tributylphosphorotrithioate, 2,3,5-tri-iodobenzoic acid, trinexapac-ethyl anduniconazole; H) herbicides selected from the group consisting ofacetamides: acetochlor, alachlor, butachlor, dimethachlor, dimethenamid,flufenacet, mefenacet, metolachlor, metazachlor, napropamide,naproanilide, pethoxaimid, pretilachlor, propachlor, thenylchlor; aminoacid derivatives: bilanafos, glufosinate, sulfosate;aryloxyphenoxypropionates: clodinafop, cyhalofop-butyl, fenoxaprop,fluazifop, haloxyfop, metamifop, propaquizafop, quizalofop,quizalo-fop-P-tefuryl; Bipyridyls: diquat, paraquat; (thio)carbamates:asulam, butylate, carbetamide, desmedipham, dimepiperate, eptam (EPTC),esprocarb, molinate, orbencarb, phenmedipham, prosulfocarb,pyributicarb, thiobencarb, triallate; cyclohexanediones: butroxydim,clethodim, cycloxydim, profoxydim, sethoxydim, tepraloxydim,tralkoxydim; dinitroanflines: benfluralin, ethalfluralin, oryzalin,pendimethalin, prodiamine, trifluralin; diphenyl ethers: acifluorfen,aclonifen, bifenox, diclofop, ethoxyfen, fomesafen, lactofen,oxyfluorfen; hydroxybenzonitriles: bomoxynil, dichlobenil, ioxynil;imidazolinones: imazamethabenz, imazamox, imazapic, imazapyr, imazaquin,imazethapyr; phenoxy acetic acids: clomeprop, 2,4-dichlorophenoxyaceticacid (2,4-D),2,4-DB, dichlorprop, MCPA, MCPA-thioethyl, MCPB, Mecoprop;pyrazines: chloridazon, flufenpyr-ethyl, fluthiacet, norflurazon,pyridate; pyridines: aminopyraild, clopyralid, diflufenican, dithiopyr,fluridone, fluroxypyr, picloram, picolinafen, thiazopyr; sulfonyl ureas:amidosulfuron, azimsulfuron, bensulfuron, chlorimuron-ethyl,chlorsulfuron, cinosulfuron, cyclosulfamuron, ethoxysulfuron,flazasulfuron, flucetosulfuron, flupyrsulfuron, foramsulfuron,halosulfuron, imazosulfuron, iodosulfuron, mesosulfuon,metsulfuron-methyl, nicosulfuron, oxasulfuron, primisulfuron,prosulfuron, pyrazosulfuron, rimsulfuron, sulfometuron, sulfosulfuron,thifensulfuron, triasulfuron, tribenuron, trifloxysulfuron,triflusulfuron, tritosulfuron,1-((2-chloro-6-propyl-imidazol[1,2-b]pyridazin-3-yl)sulfonyl)-3-(4,6-dimethoxy-pyrimidin-2-yl)urea;triazines: ametryn, atrazine, cyanazine, dimethametryn, ethiozin,hexazinone, metamitron, metribuzin, prometryn, simazine, terbuthylazine,terbutryn, triaziflam; ureas: chlorotoluron, daimuron, diuron,fluometuron, isoproturon, linuron, methabenzthiazuron, tebuthiuron;other acetolactate synthase inhibitors: bispyribac-sodium,cloransulam-methyl, diclosulam, florasulam, flucarbazone, flumetsulam,metosulam, ortho-sulfamuron, penoxsulam, propoxycarbazone,pyribambenz-propyl, pyribenzoxim, pyriftalid, pyriminobac-methyl,pyrimisulfan, pyrithiobac, pyroxasulfone, pyroxsulam; others:amicarbazone, aminotriazole, anilofos, beflubutamid, benazolin,bencarbazone, benfluresate, benzofenap, bentazone, benzobicyclon,bromacil, bromobutide, butafenacil, butamifos, cafenstrole,carfentrazone, cinidon-ethlyl, chlorthal, cinmethylin, clomazone,cumyluron, cyprosulfamide, dicamba, difenzoquat, diflufenzopyr,Drechslera monoceras, endothal, ethofumesate, etobenzanid, fentrazamide,flumiclorac-pentyl, flumioxazin, flupoxam, flurochloridone, flurtamone,indanofan, isoxaben, isoxaflutole, lenacil, propanil, propyzamide,quinclorac, quinmerac, mesotrione, methyl arsonic acid, naptalam,oxadiargyl, oxadiazon, oxaziclomefone, pentoxazone, pinoxaden,pyraclonil, pyraflufen-ethyl, pyrasulfotole, pyrazoxyfen, pyrazolynate,quinoclamine, saflufenacil, sulcotrione, sulfentrazone, terbacil,tefuryltrione, tembotrione, thiencarbazone, topramezone,4-hydroxy-3-[2-(2-methoxy-ethoxymethyl)-6-trifluoromethyl-pyridine-3-carbonyl]-bicyclo[3,2,1]oct-3-en-2-one,(3-[2-chloro-4-fluoro-5-(3-methyl-2,6-dioxo-4-trifluoromethyl-3,6-dihydro-2H-pyrimidin-1-yl)-phenoxyl]-pyridin-2-yloxy)-aceticacid ethyl ester, 6-amino-5-chloro-2-cyclopropyl-pyrimidine-4-carboxylicacid methyl ester,6-chloro-3-(2-cyclopropyl-6-methyl-phenoxy)-pyridazin-4-ol,4-amino-3-chloro-6-(4-chloro-phenyl)-5-fluoro-pyridine-2-carboxylicacid,4-amino-3-chloro-6-(4-chloro-2-fluoro-3-methoxy-phenyl)-pyridine-2-carboxylicacid methyl ester, and4-amino-3-chloro-6-(4-chloro-3-dimethylamino-2-fluoro-phenyl)-pyridine-2-carboxylicacid methyl ester; J) insecticides selected from the group consisting oforgano(thio)phosphates: acephate, azamethiphos, azinphos-methyl,chlorpyrifos, chlorpyrifos-methyl, chlorfenvinphos, diazinon,dichlorvos, dicrotophos, dimethoate, disulfoton, ethion, fenitrothion,fenthion, isoxathion, malathion, methamidophos, methidathion,methyl-parathion mevinphos, monocrolophos, oxydemeton-methyl, paraoxon,parathion, phenthoate, phosalone, phosmet, phosphamidon, phorate,phoxim, pirimiphos-methyl, profenofos, prothiofos, sulprophos,tetrachlorvinphos, terbufos, triazophos, trichlorfon; carbamates:alanycarb, aldicarb, bendiocarb, benfuracarb, carbaryl, carbofuran,carbosulfan, fenoxycarb, furathiocarb, methiocarb, methomyl, oxamyl,pirimicarb, propoxur, thiodicarb, triazamate; pyrethroids: allethrin,bifenthrin, cyfluthrin, cyhalothrin, cyphenothrin, cypermethrin,alpha-cypermethrin, beta-cypermethrin, zeta-cypermethrin, deltamethrin,esfenvalerate, etofenprox, fenpropathrin, fenvalerate, imiprothrin,lambda-cyhalothrin, permethrin, prallethrin, pyrethrin I and II,resmethrin, silafluofen, taufluvalinate, tefluthrin, tetramethrin,tralomethrin, transfluthrin, profluthrin, dimefluthrin; insect growthregulators: a) chitin synthesis inhibitors: benzoylureas:chlorfluazuron, cyramazin, diflubenzuon, flucycloxuron, flufenoxuon,hexaflumuron, lufenuron, novaluron, teflubenzuron, triflumuron;buprofezin, diofenolan, hexythiazox, etoxazole, clofentazine; b)ecdysone antagonists: halofenozide, methoxyfenozide, tebufenozide,azadirachtin; c) juvenoids: pyriproxyfen, methoprene, fenoxycarb; d)lipid biosynthesis inhibitors: spirodiclofen, spiromesifen,spirotetramat; nicotinic receptor agonists/antagonists compounds:clothianidin, dinotefuran, imidacloprid, thiamethoxam, nitenpyram,acetamiprid, thiacloprid,1-(2-chloro-thiazol-5-ylmethyl)-2-nitrimino-3,5-dimethyl-[1,3,5]triazinane;GABA antagonist compounds: endosulfan, ethiprole, fipronil, vaniliprole,pyrafluprole, pyriprole,5-amino-1-(2,6-dichloro-4-methyl-phenyl)-4-sulfinamoyl-1H-pyrazole-3-carbothioicacid amide; macrocyclic lactone insecticides: abamectin, emarnectin,milbemectin, lepimectin, spinosad, spinetoram; mitochondrial electrontransport inhibitor (METI) I acaricides: fenazaquin, pyridaben,tebufenpyrad, tolfenpyrad, flufenerim; METI II and III compounds:acequinocyl, fluacyprim, hydramethylnon: uncouplers: chlorfenapyr;oxidative phosphorylation inhibitors: cyhexatin, diafenthiuron,fenbutatin oxide, propargite; moulting disruptor compounds: cryomazine;mixed function oxidase inhibitors: piperonyl butoxide; sodium channelblockers: indoxacarb, metaflumizone; others: bendothiaz, bifenazate,cartap, flonicamid, pyridalyl, pymetrozine, sulfur, thiocyclam,flubendiamide, chlorantraniliprole, cyazypyr (HGW86), cyenopyrafen,flupyrazofos, cyflumetofen, amidoflumet, imicyafos, bistrifluron, andpyrifluquinazon. 4-6. (canceled)
 7. The method as claimed in claim 3,wherein the Bacillus subtilis strain with NRRL Accession No. B-21661 ora cell-free extract thereof, and/or a mutant of this strain or extracthaving all the identifying characteristics of the respective strain orextract and the at least one chemical compound are applied insuccession.
 8. The method as claimed in claim 3, wherein the Bacillussubtilis strain with NRRL Accession No. B-21661 or a cell-free extractthereof, and/or a mutant of this strain or extract having all theidentifying characteristics of the respective strain or extract and theat least one chemical compound are applied in a weight ratio of from100:1 to 1:100.
 9. The method as claimed in claim 3, wherein theBacillus subtilis strain with NRRL Accession No. B-21661 or a cell-freeextract thereof, and/or a mutant of this strain or extract having allthe identifying characteristics of the respective strain or extract andthe at least one chemical compound are applied simultaneously.
 10. Themethod as claimed in claim 1, wherein the seed is treated.
 11. Themethod as claimed in claim 1, wherein the application is carried out asin-furrow and/or foliar treatment.
 12. The method as claimed in claim 1,wherein a repeated application is carried out.
 13. The method as claimedin claim 3, wherein the at least one chemical compound is applied beforethe Pre-harvest interval while the Bacillus subtilis strain with NRRLAccession No. B-21661 or a cell-free extract thereof, and/or a mutant ofthis strain or extract having all the identifying characteristics of therespective strain or extract is applied during the Pre-harvest interval.14. The method as claimed in claim 1, wherein the agricultural plant isselected from soybean, corn, wheat, triticale, barley, oat, rye, rape,millet, rice, sunflower, cotton, sugar beet, pome fruit, stone fruit,citrus, banana, strawberry, blueberry, almond, grape, mango, papaya,peanut, potato, tomato, pepper, cucurbit, cucumber, melon, watermelon,garlic, onion, broccoli, carrot, cabbage, bean, dry bean, canola, pea,lentil, alfalfa, trefoil, clover, flax, elephant grass, grass, lettuce,sugarcane, tea, tobacco and coffee; each in its natural or geneticallymodified form.
 15. The use of the Bacillus subtilis strain with NRRLAccession No. B-21661 or a cell-free extract thereof, and/or a mutant ofthis strain or extract having all the identifying characteristics of therespective strain or extract for increasing the vigor and/or crop yieldof agricultural plants under essentially non-existent pathogen pressurecomprising the step of treating the plants, the plant propagules, theseed of the plants and/or the locus where the plants are growing or areintended to grow with said strain and/or extract.